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Gene/Clone	Species	Stage	Anatomy Item	Experimenter
sox2	xenopus

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Experiment details for sox2

van Grunsven LA et al. (2007) Assay

Gene	Clone	Species	Stages	Anatomy
sox2		xenopus	unspecified stage

Gene	Clone	Species	Stages	Anatomy
sox2.L		laevis	NF stage 14	ectoderm , neuroectoderm , neural plate , anterior neural fold

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Fig. 2. Effect of BMP signaling on XSip1 activity. (A–I) Animal caps derived from embryos injected at the four cell-stage (250 pg/blastomere) with Geminin RNA (A), XSip1 RNA (B, C, E–I) or XSip1δSBD mRNA (D) with or without CA-Alk3 RNA (100 pg) analyzed at neurula stage by in situ hybridization with the indicated probes. For each marker, control non-injected animal caps are shown on the left. Non-injected control embryos and CA-Alk3 RNA-injected (100 pg) embryos at neurula stage (dorsal view, anterior right) are shown on the right. Note that CA-Alk3 blocks XSip1's ability to induce the neuronal markers Sox2 and NCAM (B and C) and the repression of Gata2 expression (E). In contrast, CA-Alk3 does not affect XSip1's ability to block several other epidermal genes like epidermal keratin, TA-2, Hya-1 and Vgl-4 (E–I). (J) Lateral views of embryos injected with XSip1 RNA alone or together with CA-Alk3 RNA and stained for epidermal keratin. Co-expression of CA-Alk3 does not affect XSip1 repression of epidermal keratin. Respective inductions/inhibitions in “+CA-Alk3 caps and embryos”: (A) all positive, n = 35; (B) all inhibited, n = 18; (C) all inhibited, n = 52; (D) all inhibited, n = 36; (E) none inhibited, n = 40; (F) all inhibited, n = 22; (G) all inhibited, n = 25; (H) all inhibited, n = 28; (J) all inhibited, n = 33. Arrows in panels E–J indicate the injected area.

Gene	Clone	Species	Stages	Anatomy
sox2.L		laevis	NF stage 19	neuroectoderm , neural plate

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Fig. 6. CtBP dependence of XSip1-mediated neuralizing activity. Whole-mount in situ analysis of animal caps and embryos analyzed for expression of Sox2 or Ep. keratin. (A) Animal caps derived from embryos injected with NZF-CtBP or NZF-GFP RNA (250 pg/blastomere) analyzed for Sox2 expression and embryos injected with the same RNA analyzed for epidermal keratin expression. (B) Lateral view of NZF-CID, NZF, and NZF-CIDmut injected embryos (250 pg/blastomere) analyzed for Ep. keratin expression. (C) Animal caps derived from embryos injected with XSip1 mRNA (100 pg/blastomere) co-injected with mSip1 CID or mSip1 CIDmut RNA (1 ng/blastomere) and analyzed for Sox2 expression. Note that mSip1 CID, but not mSip1 CIDmut, reduces XSip1's ability to induce Sox2. A control Western blot for Myc-XSip1 and Myc-CID content in the animal caps is shown in the right panel. (D) XSip1 and CtBP-VP16 co-expressing animal caps showing reduced level of Sox2 expression. Non-injected controls are shown on the right and XSip1 controls are shown in panel E. (E) Animal caps derived from embryos injected with XSip1 RNA (100 pg/blastomere) alone or co-injected with XCtBP MO (20 ng/blastomere) or co-injected with the XCtBP MO and mCtBP2 (200 pg/blastomere) analyzed for Sox2 expression. Caps expressing mCtBP2 alone and embryos analyzed for Sox2 are shown as controls. The efficiency of the XCtBP MO is shown in the Western blot analysis of in vitro transcription/translation reactions of XCtBP performed in the presence of increasing amounts of XCtBP MO. (F) Animal caps derived from XSip1 injected embryos were cultured from the time of their excision in the presence or absence of 400 nM TSA until stage 14 and then processed by in situ hybridization for Sox2 expression. Note that TSA treatment inhibits the ability of XSip1 to induce Sox2. Respective inhibition/inductions (A) 70% positive caps, n = 27 for NZF-CtBP; none for NZF-GFP (n = =25); 100% inhibited embryos with NZF-CtBP (n = 30); none with NZF-GFP (n = 32); (B) 60% inhibited embryos (n = 25) for NZF-CID; none for NZF alone (n = 30) and for NZF-CIDmut (n = 32); (C) 90% with reduced staining (n = 32) for XSip1 + CID; none (n = 35) for XSip1 + CIDmut; (D) all with reduced staining (n = 40) for XSip1 + CtBP-VP16; no staining in control caps (n = 30); (E) all strongly positive (n = 38) for XSip1; all with reduced staining (n = 39) for XSip1 + MO CtBP; 50% with strongly positive, n = 30 for XSip1 + CtBP MO and mCtBP2; none positive, n = 35 for mCtBP2; (F) 85% positive caps for XSip1 (n = 38) and none for XSip1 + TSA (n = 45).