Oocytes of a single individual of Ciona intestinalis (Roscoff, France) were fertilized by a mixture of sperm of two individuals and development was followed microscopically. Embryos were snapfrozen in liquid nitrogen at the appropriate stage and kept at -80°C. Samples at stage 0, 8, 12, 15, 21, 26 were collected from the same fertilzation, while sample from stage 11 was independently collected from a different individual. Total RNA was prepared with the RNA II kit of Magerey Nagel and the quality was checked with the Bioanalyzer, all samples had a RIN value of higher than 9. Strand specific libraries were constructed and the sequencing (50PE) was performed with Illumina HiSeq 2000 technology (BGI, Hong Kong, China). Reads were aligned onto C.robusta genome and considered to reflect the expression of the C.robusta orthologous genes.

Conversion to Aniseed format of data from Yutaka Satou’s gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :Tbx6b/c/d.
Expression pattern : No signal was detected

WT Ciona robusta 2-cell stage embryo (anterior to the left) probed for Tbx6-r.b expression.
No Tbx6-r.b expression is observed.

WT Ciona robusta 4-cell stage embryo (anterior to the left) probed for Tbx6-r.b expression.
No Tbx6-r.b expression is observed.

WT Ciona robusta 8-cell stage embryo (anterior to the left) probed for Tbx6-r.b expression.
No Tbx6-r.b expression is observed.

Conversion to Aniseed format of data from Yutaka Satou’s gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :Tbx6b/c/d.
Expression pattern : no expression.

Vegetal view of a WT Ciona robusta 16-cell stage embryo probed for Tbx6-r.b expression (and injected with iin vitro-transcribed GFP:H2B control mRNA).
Tbx6-r.b expression is observed in B5.1 blastomer pair.

Vegetal view of a 16-cell stage embryo probed for Ci-Tbx6b expression. Cells where mRNA is first detected are indicated by arrowheads.

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :Tbx6b/c/d.
Expression pattern : B5.1

Vegetal pole view of a 16-cell stage embryo probed with Ci-Tbx6b expression.

Animal and vegetal views of a 16-cell stage embryo probed for Tbx6.b expression.
Original annotation: As observed in normal embryos, in embryos injected with the control MO, Tbx6.b was expressed only in the PVD.

Expression of Ci-Tbx6b was detected by whole-mount in situ hybridization. Vegetal view of a 16-cell stage embryo.

Illustrations on the left depict the constructs. The numbers indicate the relative nucleotide positions from the transcription start site of Tbx6.b. Mutated Tcf7-binding sites are indicated by X. Graphs show the percentage of blastomeres expressing the reporter in the anterior vegetal blastomeres, in the posterior vegetal blastomeres, and in the animal blastomeres. Note that not all cells or embryos could express the reporter because of mosaic incorporation of the electroporated plasmid.

Illustrations on the left depict the constructs. The numbers indicate the relative nucleotide positions from the transcription start site of Tbx6.b. Mutated Tcf7-binding sites are indicated by X. Graphs show the percentage of blastomeres expressing the reporter in the anterior vegetal blastomeres, in the posterior vegetal blastomeres, and in the animal blastomeres. Note that not all cells or embryos could express the reporter because of mosaic incorporation of the electroporated plasmid.

Illustrations on the left depict the constructs. The numbers indicate the relative nucleotide positions from the transcription start site of Tbx6.b. Mutated Tcf7-binding sites are indicated by X. Graphs show the percentage of blastomeres expressing the reporter in the anterior vegetal blastomeres, in the posterior vegetal blastomeres, and in the animal blastomeres. Note that not all cells or embryos could express the reporter because of mosaic incorporation of the electroporated plasmid.
Images showing Gfp expression in embryos electroporated with the fourth and last constructs shown in (A). Scale bar, 100 μm.
"Our analyses using a series of deletion reporter constructs identified the −189 bp upstream of Tbx6.b as a region sufficient to drive reporter expression in the PVD at the 16-cell stage"

Illustrations on the left depict the constructs. The numbers indicate the relative nucleotide positions from the transcription start site of Tbx6.b. Mutated Tcf7-binding sites are indicated by X. Graphs show the percentage of blastomeres expressing the reporter in the anterior vegetal blastomeres, in the posterior vegetal blastomeres, and in the animal blastomeres. Note that not all cells or embryos could express the reporter because of mosaic incorporation of the electroporated plasmid.

Illustrations on the left depict the constructs. The numbers indicate the relative nucleotide positions from the transcription start site of Tbx6.b. Mutated Tcf7-binding sites are indicated by X. Graphs show the percentage of blastomeres expressing the reporter in the anterior vegetal blastomeres, in the posterior vegetal blastomeres, and in the animal blastomeres. Note that not all cells or embryos could express the reporter because of mosaic incorporation of the electroporated plasmid.
Images showing Gfp expression in embryos electroporated with the fourth and last constructs shown in (A). Scale bar, 100 μm.
"When two putative Tcf7-binding sites were mutated, the expression of the reporter was completely abolished"

Illustrations on the left depict the constructs. The numbers indicate the relative nucleotide positions from the transcription start site of Tbx6.b. Mutated Tcf7-binding sites are indicated by X. Graphs show the percentage of blastomeres expressing the reporter in the anterior vegetal blastomeres, in the posterior vegetal blastomeres, and in the animal blastomeres. Note that not all cells or embryos could express the reporter because of mosaic incorporation of the electroporated plasmid.

First picture
Four repeats of the critical upstream sequence of Tbx6.b with or without mutations were ligated to the Brachyury basal promoter. Graphs show the percentage of blastomeres expressing the reporter among the anterior vegetal blastomeres and among the posterior vegetal blastomeres. A graph showing the percentage in the animal hemisphere is not shown, because no embryos expressed the reporter in the animal hemisphere. Note that not all cells or embryos could express the reporter because of mosaic incorporation of the electroporated plasmid.
Second picture
Images showing the reporter gene expression in embryos electroporated with the construct shown in the first image.
“While the basal promoter alone cannot drive reporter expression, the fusion construct was specifically expressed in the PVD. Therefore, these sequences contained sufficient elements to drive the reporter expression in the PVD.”

Four repeats of the critical upstream sequence of Tbx6.b with or without mutations were ligated to the Brachyury basal promoter. Graphs show the percentage of blastomeres expressing the reporter among the anterior vegetal blastomeres and among the posterior vegetal blastomeres. A graph showing the percentage in the animal hemisphere is not shown, because no embryos expressed the reporter in the animal hemisphere. Note that not all cells or embryos could express the reporter because of mosaic incorporation of the electroporated plasmid.
“While the basal promoter alone cannot drive reporter expression, the fusion construct was specifically expressed in the PVD. Therefore, these sequences contained sufficient elements to drive the reporter expression in the PVD.”

Four repeats of the critical upstream sequence of Tbx6.b with or without mutations were ligated to the Brachyury basal promoter. Graphs show the percentage of blastomeres expressing the reporter among the anterior vegetal blastomeres and among the posterior vegetal blastomeres. A graph showing the percentage in the animal hemisphere is not shown, because no embryos expressed the reporter in the animal hemisphere. Note that not all cells or embryos could express the reporter because of mosaic incorporation of the electroporated plasmid.
“While the basal promoter alone cannot drive reporter expression, the fusion construct was specifically expressed in the PVD. Therefore, these sequences contained sufficient elements to drive the reporter expression in the PVD.”

Four repeats of the critical upstream sequence of Tbx6.b with or without mutations were ligated to the Brachyury basal promoter. Graphs show the percentage of blastomeres expressing the reporter among the anterior vegetal blastomeres and among the posterior vegetal blastomeres. A graph showing the percentage in the animal hemisphere is not shown, because no embryos expressed the reporter in the animal hemisphere. Note that not all cells or embryos could express the reporter because of mosaic incorporation of the electroporated plasmid.
Second picture
Images showing the reporter gene expression in embryos electroporated with the construct shown in the first image.
“While the basal promoter alone cannot drive reporter expression, the fusion construct was specifically expressed in the PVD. Therefore, these sequences contained sufficient elements to drive the reporter expression in the PVD.”
“The mutant construct μC induced ectopic expression of the reporter in the AVD. Therefore, the region mutated in μC (region C) is likely to repress expression in the AVD”

Four repeats of the critical upstream sequence of Tbx6.b with or without mutations were ligated to the Brachyury basal promoter. Graphs show the percentage of blastomeres expressing the reporter among the anterior vegetal blastomeres and among the posterior vegetal blastomeres. A graph showing the percentage in the animal hemisphere is not shown, because no embryos expressed the reporter in the animal hemisphere. Note that not all cells or embryos could express the reporter because of mosaic incorporation of the electroporated plasmid.
“While the basal promoter alone cannot drive reporter expression, the fusion construct was specifically expressed in the PVD. Therefore, these sequences contained sufficient elements to drive the reporter expression in the PVD.”

Four repeats of the critical upstream sequence of Tbx6.b with or without mutations were ligated to the Brachyury basal promoter. Graphs show the percentage of blastomeres expressing the reporter among the anterior vegetal blastomeres and among the posterior vegetal blastomeres. A graph showing the percentage in the animal hemisphere is not shown, because no embryos expressed the reporter in the animal hemisphere. Note that not all cells or embryos could express the reporter because of mosaic incorporation of the electroporated plasmid.
“While the basal promoter alone cannot drive reporter expression, the fusion construct was specifically expressed in the PVD. Therefore, these sequences contained sufficient elements to drive the reporter expression in the PVD.”

Four repeats of the critical upstream sequence of Tbx6.b with or without mutations were ligated to the Brachyury basal promoter. Graphs show the percentage of blastomeres expressing the reporter among the anterior vegetal blastomeres and among the posterior vegetal blastomeres. A graph showing the percentage in the animal hemisphere is not shown, because no embryos expressed the reporter in the animal hemisphere. Note that not all cells or embryos could express the reporter because of mosaic incorporation of the electroporated plasmid.
“While the basal promoter alone cannot drive reporter expression, the fusion construct was specifically expressed in the PVD. Therefore, these sequences contained sufficient elements to drive the reporter expression in the PVD.”

Illustrations on the left depict the constructs. A potential repressive element in the upstream sequence of Tbx6.b was inserted into the upstream sequences of Fgf9/16/20. Graphs on the right show the percentage of blastomeres expressing the reporter gene in the anterior vegetal blastomeres and in the posterior vegetal blastomeres. A graph showing the percentage in the animal hemisphere is not shown, because no embryos expressed the reporter in the animal hemisphere.
"With this insertion, the construct rarely drove reporter expression in the AVD"
"Although the result does not necessarily indicate that the 22 bp region is the only repressive element in the upstream region of Tbx6.b, it showed that this 22 bp region has a repressive activity."

Constructs depicted in the illustrations on the left were injected with or without an MO against Zic-r.a. The green boxes indicate the Gfp reporter gene and SV40 polyadenylation signal. Graphs on the right show the percentage of blastomeres expressing the reporter gene in the anterior vegetal blastomeres and in the posterior vegetal blastomeres.

First picture
Constructs depicted in the illustrations on the left were injected with or without an MO against Zic-r.a. Graphs on the right show the percentage of blastomeres expressing the reporter gene in the anterior vegetal blastomeres and in the posterior vegetal blastomeres.
Second picture
Photographs of embryo injected with the Fgf9/16/20 reporter construct with the repressive element of Tbx6.b. Scale bar, 100 μm.

Vegetal view of a 32-cell stage embryo probed for Ci-Tbx6b expression.

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :Tbx6b/c/d.
Expression pattern : B6.2, B6.4
Comment : The expression in the B6.2 is dispersed within cytoplasm and looked weak.

Vegetal view of a late 32-cell stage embryo.Tbx6b expression was detected in B6.1, B6.2, B6.4 blastomeres (signal in B6.1 and B6.2 is scarcely visible).

Vegetal pole view of a late 32-cell stage embryo probed with Ci-Tbx6b expression.

Expression of Ci-Tbx6b was detected by whole-mount in situ hybridization. Vegetal view of a late 32-cell stage embryo.

32-cell stage embryos electroporated with the 2.4 kb wild-type Ci-Tbx6b CRM, region upstream of Ci-Tbx6b from -2339bp to -13bp.
The first picture is a schematic representation of the electroporated region. Green boxes represent putative Ci-Macho1-binding sites; black arrows indicate their orientations. The orange rectangle depicts the proximal region of the 2.4 kb fragment.
The percentages of transgenic embryos displaying the patterns shown, and the total number of embryos scored, are indicated at the bottom of each panel.

Original Annotation : A strong hybridization signal was detected in the B6.4 pair of muscle precursors.

32-cell stage embryos electroporated with the 2.4 kb CRM with mutations in all Ci-Macho1-binding sites.
The first picture is a schematic representation of the electroporated region. Green boxes represent putative Ci-Macho1-binding sites; black arrows indicate their orientations. The orange rectangle depicts the proximal region of the 2.4 kb fragment.
The percentages of transgenic embryos displaying the patterns shown, and the total number of embryos scored, are indicated at the bottom of each panel.

Original Annotation : No expression of the reporter at the 32-cell stage.

32-cell stage embryos electroporated with the 862 bp Ci-Tbx6b CRM, which does not contain the Ci-Macho1-binding sites.
The first picture is a schematic representation of the electroporated region. The orange rectangle depicts the proximal region of the 2.4 kb fragment.
The percentages of transgenic embryos displaying the patterns shown, and the total number of embryos scored, are indicated at the bottom of each panel.

Original Annotation : No detectable signal at the 32-cell stage.

32-cell stage embryos electroporated with the fragment 266bp Ci-Tbx6b CRM containing the region upstream of Ci-Tbx6b from -145bp to -13bp upstream of basal promoter of Ci-Tbx6b.
The first picture is a schematic representation of the electroporated region.
The 266 bp proximal region of the Ci-Tbx6b CRM contains putative binding sites for CREB (blue diamond), T-box (red oval) and bHLH ("AC"-core E-box; pink triangle) transcription factors.
The percentages of transgenic embryos displaying the patterns shown, and the total number of embryos scored, are indicated at the bottom of each panel.

Original Annotation : The majority of the embryos electroporated with the 266 bp construct did not show staining at the 32-cell stage.

Embryos electroporated with either the distal muscle CRM and fixed at the 32-cell stage.
The distal region contains two E-boxes (pink triangles).
The percentages of transgenic embryos displaying the patterns shown, and the total number of embryos scored, are indicated at the bottom of each panel.

No reporter expression was detected.

Vegetal view of a 64-cell stage wild type embryo, showing expression of Ci-Tbx6b.

According to the picture, Ci-Tbx6b was expressed in B7.7, B7.5, B7.4 and B.8 cell pair.

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :Tbx6b/c/d.
Expression pattern : B7.3, B7.4, B7.5, B7.7, B7.8.
Comment : The expression in the B7.3 and B7.4 is dispersed within cytoplasm and looked weak.

Vegetal pole view of a 64-cell stage embryo probed with Ci-Tbx6b expression.

Expression of Ci-Tbx6b was detected by whole-mount in situ hybridization. Vegetal view of a 64-cell stage embryo.

Vegetal view of 110-cell stage wild type embryo probed for Tbx6b expression.

Vegetal view of a 112-cell stage embryo probed for Ci-Tbx6b expression.

112-cell stage wild type embryos probed for Ci- Tbx6b expression.
In the second picture, the notochord and its precursors are outlined by a dashed red line.

Original Annotation : Ci-Tbx6b is expressed at the 112-cell stage in muscle precursors (orange arrowheads).

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :Tbx6b/c/d.
Expression pattern : B8.7, B8.8, B7.5, B8.15, B8.16.
Comment : The expression in the B8.15 and B8.16 is dispersed within cytoplasm and looked weak.

Vegetal pole view of a 110-cell stage embryo probed with Ci-Tbx6b expression.

Expression of Ci-Tbx6b was detected by whole-mount in situ hybridization. Vegetal view of a 110-cell stage embryo.

Aniseed annotation:
According to the picture and to the others data stored in ANISEED, it seems that there is also expression in B7.5 and B8.16 cell pairs.

112-cell stage embryos electroporated with the 2.4 kb wild-type Ci-Tbx6b CRM, region upstream of Ci-Tbx6b from -2339bp to -13bp.
The first picture is a schematic representation of the electroporated region. Green boxes represent putative Ci-Macho1-binding sites; black arrows indicate their orientations. The orange rectangle depicts the proximal region of the 2.4 kb fragment.
The percentages of transgenic embryos displaying the patterns shown, and the total number of embryos scored, are indicated at the bottom of each panel.

Original Annotation : At the 112-cell stage, lacZ expression was detected in descendants of B6.4 blastomeres and in the B8.7 and B8.8 pairs, which derive from the B6.2 pair.

112-cell stage embryos electroporated with the 2.4 kb CRM with mutations in all Ci-Macho1-binding sites.
The first picture is a schematic representation of the electroporated region. Green boxes represent putative Ci-Macho1-binding sites; black arrows indicate their orientations. The orange rectangle depicts the proximal region of the 2.4 kb fragment.
The percentages of transgenic embryos displaying the patterns shown, and the total number of embryos scored, are indicated at the bottom of each panel.

Original Annotation : No expression of the reporter at the 112-cell stage.

112-cell stage embryos electroporated with the 862 bp Ci-Tbx6b CRM, which does not contain the Ci-Macho1-binding sites.
The first picture is a schematic representation of the electroporated region. The orange rectangle depicts the proximal region of the 2.4 kb fragment.
The percentages of transgenic embryos displaying the patterns shown, and the total number of embryos scored, are indicated at the bottom of each panel.

Original Annotation : No detectable signal at the 112-cell stage.

112-cell stage embryos electroporated with the fragment 266bp Ci-Tbx6b CRM containing the region upstream of Ci-Tbx6b from -145bp to -13bp upstream of basal promoter of Ci-Tbx6b.
The first picture is a schematic representation of the electroporated region.
The 266 bp proximal region of the Ci-Tbx6b CRM contains putative binding sites for CREB (blue diamond), T-box (red oval) and bHLH ("AC"-core E-box; pink triangle) transcription factors.
The percentages of transgenic embryos displaying the patterns shown, and the total number of embryos scored, are indicated at the bottom of each panel.

Original Annotation : The majority of the embryos electroporated with the 266 bp construct did not show staining at the 112-cell stage.

Embryos electroporated with either the distal muscle CRM and fixed at the 112-cell stage.
The distal region contains two E-boxes .

Original Annotation : In the majority of the embryos analyzed, high levels of lacZ transcripts were predominantly detected in only one pair of muscle precursors although a few embryos also showed a faint signal in additional muscle precursors (orange arrowheads), including the mixed-lineage A8.16 blastomeres (orange and blue arrowhead).

Vegetal view of a gastrula stage embryo probed for Ci-Tbx6b expression.

Late gastrula embryos probed for Ci-Tbx6b expression.
The notochord and its precursors are outlined by a dashed red line.

Original Annotation : Wild-type embryos show expression of Ci-Tbx6b in muscle precursors at the gastrula stage.

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :Tbx6b/c/d.
Expression pattern : muscle

Expression of Ci-Tbx6b was detected by whole-mount in situ hybridization. View of a late gastrula stage embryo.

Vegetal pole view of a late tailbud stage wild type embryo probed for Ci-Tbx6b expression. This picture shows expression in the rowI/column4 (A9.31) an din the tail muscles. Numbers indicate any expression over the total number of embryos analysed. Nuclei are labeled with Hoechst (white) to allow the easy identification of individual cells.

Vegetal view of a late gastrula wild type embryo probed for Ci-tbx6b expression.

Original Annotation : At this stage the Ci-tbx6b staining is detected in the rowI column 4 (A9.31) and in the tail muscle.

Early neurula embryos electroporated with the 2.4 kb CRM with mutations in all Ci-Macho1-binding sites.
The first picture is a schematic representation of the electroporated region. Green boxes represent putative Ci-Macho1-binding sites; black arrows indicate their orientations. The orange rectangle depicts the proximal region of the 2.4 kb fragment.
The percentages of transgenic embryos displaying the patterns shown, and the total number of embryos scored, are indicated at the bottom of each panel.

Original Annotation : The vast majority of embryos carrying the 2.4 kb CRM with mutations in all Ci-Macho1-binding sites displayed active transcription in the posterior-most muscle cells only at the neurula stage.

Early neurula embryos electroporated with the 862 bp Ci-Tbx6b CRM, which does not contain the Ci-Macho1-binding sites.
The first picture is a schematic representation of the electroporated region. The orange rectangle depicts the proximal region of the 2.4 kb fragment.
The percentages of transgenic embryos displaying the patterns shown, and the total number of embryos scored, are indicated at the bottom of each panel.

Original Annotation : LacZ expression became detectable by the early neurula stage in only the posterior-most muscle cells (note that the asymmetrical expression is due to mosaic incorporation of the transgene).

Early neurula stage embryos electroporated with the fragment 266bp Ci-Tbx6b CRM containing the region upstream of Ci-Tbx6b from -145bp to -13bp upstream of basal promoter of Ci-Tbx6b.
The first picture is a schematic representation of the electroporated region.
The 266 bp proximal region of the Ci-Tbx6b CRM contains putative binding sites for CREB (blue diamond), T-box (red oval) and bHLH ("AC"-core E-box; pink triangle) transcription factors.
The percentages of transgenic embryos displaying the patterns shown, and the total number of embryos scored, are indicated at the bottom of each panel.

Original Annotation : By the early neurula stage, 87% of the embryos showed strong staining in muscle cells.

Embryos electroporated with either the distal muscle CRM and fixed at the early neurula stage.
The distal region contains two E-boxes .

Original Annotation : Consistent with this early pattern, at the early neurula stage beta-galactosidase accumulation was detected only in a small subset of muscle and mesenchyme precursors.

Early neurula embryos electroporated with the 2.4 kb wild-type Ci-Tbx6b CRM, region upstream of Ci-Tbx6b from -2339bp to -13bp.
The first picture is a schematic representation of the electroporated region. Green boxes represent putative Ci-Macho1-binding sites; black arrows indicate their orientations. The orange rectangle depicts the proximal region of the 2.4 kb fragment.
The percentages of transgenic embryos displaying the patterns shown, and the total number of embryos scored, are indicated at the bottom of each panel.

Original Annotation : In early neurulae, lacZ expression was detected only in the posterior-most muscle cells. This pattern faithfully recapitulates the expression of Ci-Tbx6b.

Dorsal view of a neurula stage embryo probed for Ci-Tbx6b expression.

Conversion to Aniseed format of data from Yutaka Satous gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :Tbx6b/c/d.
Expression pattern : muscle.
Comment : The primary muscle was stained lightly and the secondary muscle was stained deeply.

Expression of Ci-Tbx6b was detected by whole-mount in situ hybridization. View of a mid-neurula stage embryo.

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :Tbx6b/c/d.
Expression pattern : No distinct zygotic signal was detected

Embryos electroporated with either the distal muscle CRM and fixed at the early tailbud stage.
The distal region contains two E-boxes .

Original Annotation : By the early tailbud stage, the activity of the CRM had considerably expanded, to encompass virtually all muscle cells (orange arrowheads), as well as part of the nerve cord (blue arrowheads) and mesenchyme cells (pink arrowheads).

Mid tailbud embryos probed for Ci-Tbx6b expression. Embryos are oriented with anterior to the left, dorsal up.
In the second picture, the notochord and its precursors are outlined by a dashed red line.

Original Annotation : Ci-Tbx6b is expressed at the 112-cell stage in muscle precursors but is no longer expressed by the early tailbud stage.

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :Tbx6b/c/d.
Expression pattern : No distinct zygotic signal was detected

A 112 bp subfragment of the 266 bp CRM, containing the three putative binding sites(CREB (blue diamond), T-box (red oval) and E-box (pink triangle)) was cloned upstream of the Ci-FoxA-a basal promoter (pink box).
Inset show transgenic embryos fixed at the neurula stage.
The number of embryos scored for each construct is shown in white.
Low-magnification image of mid-tailbud embryos electroporated with the wild-type 112 bp Ci-Tbx6b CRM.

Aniseed Annotation : The reporter was expressed in the muscle and mesenchyme.

A 112 bp subfragment of the 266 bp CRM, containing the three putative binding sites(CREB (blue diamond), T-box (red oval) and E-box (pink triangle)) was cloned upstream of the Ci-FoxA-a basal promoter (pink box). Here the T-box site was mutated (white crossed oval).
Inset show transgenic embryos fixed at the neurula stage.
The number of embryos scored for each construct is shown in white.

Low-magnification image of mid-tailbud embryos electroporated with the wild-type 112 bp Ci-Tbx6b CRM.

Original Annotation : When the T-box binding site was mutated, a considerable reduction was observed not only in the intensity of the muscle staining but also in the percentage of embryos showing activation of the reporter gene.

Microphotographs of X-Gal-stained Ciona embryos electroporated with the 400 bp region, cloned upstream of the FoxA-a basal promoter (pink box in the schematics). The distal region contains two E-boxes (pink triangles).

Reporter expression was detected in muscle (orange arrowheads) and in the CNS (blue arrowheads).

Embryos electroporated with either the distal muscle CRM and fixed at the mid tailbud stage.
The distal region contains two E-boxes .

Original Annotation : The CRM remained active in most muscle cells (orange arrowheads) through the mid-tailbud stage.

Early neurula embryos electroporated with the 2.4 kb wild-type Ci-Tbx6b CRM, region upstream of Ci-Tbx6b from -2339bp to -13bp.
The first picture is a schematic representation of the electroporated region. Green boxes represent putative Ci-Macho1-binding sites; black arrows indicate their orientations. The orange rectangle depicts the proximal region of the 2.4 kb fragment.
The second picture is a low-magnification microphotographs of Ciona intestinalis embryos electroporated with either the 2.4 kb Ci-Tbx6b CRM.

Original Annotation : Orange and purple arrowheads indicate muscle and mesenchyme cells, respectively.

Embryos electroporated with a fragment of 456bp (named 504bp in the paper) upstream of Ci-Tbx6b, cloned upstream of Ci-FoxAa, and probed for reporter expression in muscle.
Red dashed lines represent the minimal 112bp sequence driving muscle expression.

Expression was detected in muscle.