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Figure S2. Related to Figure 2. Sox5 misexpression and knockdown causes gene expression
defects in gastrula stage embryos.
(A) phospho-Histone H3 staining of stage 15 embryos injected with Sox5 mRNA or Sox5
morpholino. No differences in proliferation are seen. (B) TUNEL staining stage 15 embryos
injected with Sox5 mRNA or Sox5 morpholino. No differences in apoptosis are seen. DNMT3bmediated
apoptosis serves as a positive control. Asterisk denotes side with β-gal staining (red)
lineage tracer. (C) In situ hybridization examining expression of MyoD in stage 15 embryos
injected with Sox5 mRNA or Sox5 morpholino in micromeres at the 8-cell stage targeting for
ectodermal tissue that will give rise to neural crest. No defects in mesoderm are noted. (D) In situ
hybridization examining expression of MyoD in stage 15 embryos injected with Sox5 mRNA or
Sox5 morpholino in macromeres at the 8 cell stage targeting for mesoderm. Mesoderm formation
as assayed by MyoD expression is perturbed, reflecting a role for Sox5 in this tissue. (E) In situ
hybridization examining expression of Sox8 in embryos injected with Sox5 mRNA. Embryos
were collected at both stage 17 and stage 23. (F) In situ hybridization examining expression of
FoxD3 and Snail2 in embryos injected with Sox5* (a shorter Sox5 isoform/splice variant
expressed in laevis). Embryos were collected at mid-neurula stages, stage 17. (G) In situ
hybridization examining expression of EPK and Sox3 in embryos injected with Sox5*. Embryos
were collected at early-neurula stages, stage 14. (H) In situ hybridization examining expression
of Msx1, Pax3 and Zic1 in embryos injected with Sox5*. Embryos were collected at earlyneurula
stages, stage 14. Sox5* behaved indistinguishably to Sox5 in these assays. (I-K) In situ
hybridization examining expression of Id3 (I), AP2 (J) and Pax3 (K) at gastrula stages (stage
11.5) in embryos injected with Sox5 or Sox5MO. Red arrow denotes blastopore. Asterisk
denotes injected side. |
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Figure 1.
Sox5 Is Required for NC and NPB Formation
(A) In situ hybridization examining FoxD3, Snail2, Sox9, and Sox10 in Xenopus embryos injected with Sox5 MO or Sox5 mRNA. Embryos were collected at midneurula stages (stage 17). Asterisk denotes injected side with β-galactosidase (β-gal) staining (red) as lineage tracer.
(B) Explant assay examining Snail2 and Sox9 in Wnt8/Chordin-induced explants that were injected with Sox5 MO. Explants were collected alongside sibling embryos cultured to midneurula stages (stage 17).
(C) In situ hybridization examining Six1 and Foxi1c in embryos injected with Sox5 MO or Sox5 mRNA. Embryos were collected at early neurula stages (stage 13).
(D) In situ hybridization examining Msx1, Pax3, and Zic1 in embryos injected with Sox5 MO or Sox5 mRNA. Embryos were collected at early neurula stages (stage 13/14).
(E) Explant assay examining Msx1, Pax3, and Zic1 in Wnt8/Chordin-induced explants that were injected with Sox5 MO. Explants were collected alongside sibling embryos cultured until early neurula stages (stage 14). Asterisk denotes injected side with β-gal staining (red) as lineage tracer.
See also Figure S1. |
