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 :GATA-a.
Expression pattern : Unlocalized signal was detected

First vegetal view and second animal view of 16-cell stage embryos injected with a full length GATAa cDNA (AK114877) GFP was fused in frame at the C-terminus of GATAa. Both UTRs in the construct come from GATAa. At the 16-cell stage GATAa protein is ubiquitously present in the embryo.

"Gata.a proteins were observed equally in all nuclei of both of the AD and VD"

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 :GATA-a.
Expression pattern : No distinct zygotic signal was detected.
Comment : Maternal signal were observed ubiquitously mainly in the early claeving stages.

First vegetal view and second animal view of a late 32-cell stage embryos injected with a full length GATAa cDNA (AK114877) containing the endogenous 3 and 5 UTR. The GATAa open reading frame was fused in frame at its C-terminus to GFP. At the 32-cell stage GATAa protein is present only in animal cells.

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 :GATA-a.
Expression pattern : No distinct zygotic signal was detected.
Comment : Maternal signal were observed ubiquitously mainly in the early claeving stages.

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 :GATA-a.
Expression pattern : No distinct zygotic signal was detected.
Comment : Maternal signal were observed ubiquitously mainly in the early claeving stages.

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 :GATA-a.
Expression pattern : No distinct zygotic signal was detected

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 :GATA-a.
Expression pattern : No distinct zygotic signal was detected.

Aniseed annotation :
week signal in the center of the embryo. Head endoderm?

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 :GATA-a.
Expression pattern : endoderm

Ventral view of a late neurula stage embryo probed for GATAa (green) expression. Nuclei of TVCs and ATMs are marked by β-gal antibody staining (red). Inset panels display magnified views of the B7.5 lineage. Note that the TVCs are the more anterior lineage, as marked by arrowheads in the first column.

Original annotation:
GATAa did not appear to be expressed in the TVCs although they were expressed in neighboring lineages.

Early-tailbud embryos electroporated with Mesp > lacZ, detected by immunofluorescence (red), and hybridized with the GATAa antisense RNA probes (green).

Original annotation:
They observed conspicuous TVC specific expression of GATAa and Tolloid.

Lateral view of an early tailbud stage embryo probed for GATAa (green) expression. Nuclei of TVCs and ATMs are marked by β-gal antibody staining (red). Inset panels display magnified views of the B7.5 lineage. Note that the TVCs are the more anterior lineage, as marked by arrowheads in the first column.

Original annotation:
Between stages 18 and 19 (9 hpf), we began to detect robust expression of GATAa in the TVCs along with continued expression in the posterior endoderm and variable, weak expression in the anterior endoderm .

Lateral view of an early tailbud stage wild type embryo probed for GATA-a 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 :GATA-a.
Expression pattern : endoderm.

Aniseed annotation :
some unidentified cells above the posterior head endoderm. Neural tissue in the neck or visceral ganglion, or some endoderm? The staining is restricted to only part of the head endoderm.

Early tailbud stage wild type embryo probed for Ci-Gataa expression. This picture shows expression in the head endoderm.

Lateral view of a late tailbud stage embryo probed for GATAa (green) expression. Nuclei of TVCs and ATMs are marked by β-gal antibody staining (red). Inset panels display magnified views of the B7.5 lineage. Note that the TVCs are the more anterior lineage, as marked by arrowheads in the first column.

Original annotation:
As the TVCs completed midline convergence, GATAa expression persisted in the TVCs.

Lateral view of a mid tailbud stage embryo co-electroporated with Mesp-lacZ (red) and Hl-GATAa-DBD and probed for GATAa (green) expression. The construct Hl-GATAa-DBD was included in embryos for a more rigourous control : this construct contains the GATAa DNA binding domain alone which is expressed in the TVCs.

Original annotation:
In Hl-GATAa-DBD control embryos, GATAa was consistently expressed in the trunk ventral cells.

Lateral view of a mid tailbud stage embryo co-electroporated with Mesp-lacZ (red) and Ttf-GATAa-DBD and probed for GATAa (green) expression. The construct Ttf-GATAa-DBD was used as a control.

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 :GATA-a.
Expression pattern : endoderm.

Aniseed annotation :
The staining is restricted to only part of the head endoderm.

Sagital view of WT mid tailbud embryo showing expression of GATAa by fluorescent ISH (green), and the position of the B7.5 lineage (red) marked by immunohistochemistry following mesp::NLS-LacZ electroporation. Expression of GATAa is detected in the postrior trunk endoderm and in the TVCs.

Lateral view of an early tailbud stage embryo probed for GATAa (green) expression. Nuclei of TVCs and ATMs are marked by β-gal antibody staining (red). Inset panels display magnified views of the B7.5 lineage. Note that the TVCs are the more anterior lineage, as marked by arrowheads in the first column.

Original annotation:
GATAa expression domains were unchanged.

Immunostaining of (A) Gata.a with specific antibodies. Images are Z-projected image stacks overlaid in pseudocolor.
(A) Gata.a mRNA is expressed in endodermal cells at this stage, and Gata.a protein was detected in nuclei of endodermal cells.
Coding sequences of Gata.a, Tcf7, and Zic-r.a were cloned into pET16b (Novagen), and Histagged proteins were produced in E. coli. After purification with NiNTA resin (Qiagen), recombinant proteins were used for immunizing rabbits and polyclonal antibodies were obtained. To detect protein localization, embryos were fixed with 3.7% formaldehyde in PBS, treated with 3% H2O2 for 30 minutes, and then incubated with the antibodies in Can-Get-Signal- Immunostain Solution B (Toyobo). The signal was visualized with a TSA kit (Invitrogen) using horseradish peroxidase-conjugated goat anti-rabbit IgG and Alexa Fluor 488 tyramide. Control embryos incubated without primary antibodies yielded no signal.