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Figure 2.
Misregulating Sox5 Phenocopies Loss of BMP Signaling
(A and B) In situ hybridization examining Sox3 (A) and EPK (B) in embryos injected with Sox5 MO or Sox5 mRNA. Embryos were collected at early neurula stages (stage 13).
(C and D) Explant assay examining Sox3 (C) and EPK (D) in explants that were either injected with Sox5 MO or Sox5 mRNA. Explants were collected alongside sibling embryos cultured until early neurula stages (stage 13).
(E and F) In situ hybridization examining Sox3 (E) and EPK (F) in embryos injected with Chordin mRNA showing a similar loss of EPK and expanded expression of Sox3. Embryos were collected at early neurula stages (stage 13/14). Asterisk denotes injected side with β-gal (red) as lineage tracer.
(G) In situ hybridization examining AP2, Id3, and Vent2 at blastula stages. Embryos were injected with Chordin, Smad7, or Sox5 MO and collected at stage 9. Asterisk denotes injected side with β-gal staining (red) as lineage tracer. See also Figure S2. |
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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. |