Figure 2.

POUV Depletion Phenotypes in Xenopus Ectoderm and EpiSCs

(A) POUV morphant ectodermal explants exhibit adhesion defects. Animal caps were excised from stage 8 blastulae injected with Control-MO or PVD2 and cultured until either late gastrula or neurula stage (stage 14). PVD2 morphant explants were slow to round up and never properly healed; by the completion of neurulation, PVD2 explants completely disaggregated.

(B) Diagram of the Oct4^LoxP/LoxP-EpiSC lines. These cells possess two conditional Pou5f1 alleles and express an inducible Cre recombinase. Addition of tamoxifen induces Cre-mediated Oct4 deletion.

(C) Immunoblot of samples from Oct4^LoxP/LoxP and Oct4^(−/−)-EpiSC 12 and 24 hr after tamoxifen treatment (0.1 μM). The bottom panel shows quantified chemiluminescent signals normalized to tubulin and relative to untreated EpiSCs. Pr-Ecad is the precursor of E-cadherin, indicated by the starred arrow.

(D) qRT-PCR of Oct4^LoxP/LoxP-EpiSCs 12 and 24 hr after tamoxifen addition. Expression was normalized to TBP, relative to untreated EpiSCs. Bars show mean ± SD.

(E) Immunofluorescence of Cre-negative (Cre-ve) and Cre-positive (Cre+ve) Oct4^LoxP/LoxP-EpiSCs 48 hr after tamoxifen treatment. The scale bar represents 50 μm.

See also Figure S2.