Xbp1 and Brachyury establish an evolutionarily conserved subcircuit of the notochord gene regulatory network
- Department of Molecular Pathobiology, New York University College of Dentistry, United States
Figures
Figure 1
Xbp1 notochord expression is dependent upon Ci-Bra.
(A) Whole-mount in situ hybridization (WMISH) of a wild-type Ciona robusta embryo at the mid-tailbud II stage (Hotta et al., 2007) with a fluorescent antisense RNA probe specific for Cr-Xbp1. …
Figure 2 with 1 supplement
Functional analysis of Ciona Xbp1.
Microphotographs of late-tailbud II (A–C), mid-tailbud I (E, F), and late-tailbud I (G–I’) C. robusta transgenic embryos, electroporated at the 1-cell stage with 50 µg of each of the plasmids …
Figure 2—figure supplement 1
Additional images of Xbp1DBD and Xbp1DBD::VP16 transgenic embryos.
Confocal microphotographs of Ciona embryos electroporated with the transgenes indicated in each panel. Anterior is to the left. (A–B’) Transgenic embryos displaying 100% incorporation of the …
Figure 3 with 3 supplements
Expression patterns of putative Cr-Xbp1 notochord targets.
(A–X) Whole-mount in situ hybridization (WMISH) of Ciona embryos ranging from early gastrula to late tailbud for the genes reported on top of each panel, with digoxigenin-labeled antisense RNA …
Figure 3—figure supplement 1
Additional expression patterns of putative Cr-Xbp1 notochord targets.
(A-J) Whole-mount in situ hybridization (WMISH) of late gastrula to late-tailbud Ciona robusta embryos for the genes reported on top of each panel, carried out with digoxigenin-labeled antisense RNA …
Figure 3—figure supplement 2
Ciona-/ascidian-specific notochord genes regulated by Xbp1.
(A–O) Whole-mount in situ hybridization (WMISH) of Ciona robusta embryos ranging from early gastrula (inset in B) to mid-tailbud II (top inset in N) for the genes reported on top of each panel, …
Figure 3—figure supplement 3
Putative Xbp1 targets expressed in tissues other than the notochord.
Whole-mount in situ hybridization (WMISH) of Ciona robusta embryos ranging from 76 cell (inset in D) to mid-tailbud II (H) for the genes reported on top of each panel, carried out with antisense RNA …
Figure 4 with 2 supplements
Effects of Cr-Xbp1 transgenes on the expression of Ci-Bra and fibrillin.
Mid-tailbud I C. robusta embryos electroporated with the transgenes reported in green font and hybridized in situ with fluorescent antisense RNA probes (red font). (A, B) Bra>GFP transgenic embryo, …
Figure 4—video 1
X-projection of confocal images of the embryo in Figure 4C.
Figure 4—video 2
X-projection of confocal images of the embryo in Figure 4F.
Figure 5 with 1 supplement
Cross-regulation between Brachyury and Xbp1 in the dorsal mesoderm and developing notochord of Xenopus.
(A–F’) Xenopus laevis embryos at the late gastrula stage (NF stage 12), control (A, C), and morphants (B, B’, D, D’, E, E’, F, F’). Injection of the Xbra translation-blocking MO (XbraMO; Shi et al., …
Figure 5—figure supplement 1
Evaluation of the effects of splice-blocking morpholino oligonucleotides on Xbra and Xbp1 transcripts.
(A, B) Schematic representation of the Xbra (A) and Xbp1 (B) gene structure. Exons (abbreviated as E) are depicted as colored boxes. For each gene, the target site of the splice-blocking MOs …
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Figure 5—figure supplement 1—source data 1
Original images of the unedited gels presented in Figure 5—figure supplement 1.
- https://cdn.elifesciences.org/articles/73992/elife-73992-fig5-figsupp1-data1-v2.pdf
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Figure 5—figure supplement 1—source data 2
Original images of the uncropped gels presented in Figure 5—figure supplement 1 with the relevant bands labeled.
- https://cdn.elifesciences.org/articles/73992/elife-73992-fig5-figsupp1-data2-v2.pdf
Figure 6 with 1 supplement
Xbra and Xbp1 morpholino-mediated knockdowns result in axis truncation.
Xbra (XbraMO) and Xbp1 (Xbp1MO) knockdowns cause comparable anterior-posterior axis elongation defects and posterior truncations. (A–C’) Xenopus embryos at NF stage 35/36. (A) Control embryo. (B, C) …
Figure 6—figure supplement 1
Notochord formation in Xbra and Xbp1 morphant embryos monitored through the expression of Shh.
Xenopus embryos at NF stage 35/36. (A) Control embryo. (B–C’) Morphant embryos injected with Xbp1MO (B, B’) and XbraMO (C, C’), displaying either mild (B, C) or severe (B’, C’) notochord phenotypes. …
Additional files
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Supplementary file 1
Ciona Xbp1 putative target genes expressed in the notochord.
- https://cdn.elifesciences.org/articles/73992/elife-73992-supp1-v2.xlsx
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Supplementary file 2
Ciona Xbp1 putative target genes expressed in tissues other than the notochord.
- https://cdn.elifesciences.org/articles/73992/elife-73992-supp2-v2.xlsx
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Supplementary file 3
Expression of Xenopus orthologs of Ciona Xbp1 notochord target genes.
- https://cdn.elifesciences.org/articles/73992/elife-73992-supp3-v2.xlsx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/73992/elife-73992-transrepform1-v2.pdf
