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Fig. 8. ROCK activity is required for epiblast differentiation and polarization. Confocal microscopy. Green, phalloidin staining detecting F-actin. Red, specific protein signal. (A) Wild-type (untreated) BM is detected by antibody to laminin {alpha}1. There is F-actin accumulation in the apical domain of the epiblast. (B) Y-27632 treatment, note the lack of columnar ectoderm and partial cavitation without F-actin accumulation. BM detected by laminin {alpha}1 antibody. (C) H-1152 treatment, antibody to collagen IV, (D) EB formed by ES cells expressing dnROCKII. Note flat internal cells without significant F-actin accumulation in the apical domain. (E,F) ZO-1 staining. The pattern is similar in the wild-type endoderm (E) and in H-1152-treated EB (F). (G,H) ROCKII staining. (G) Wild type; (H) EB formed by dnFgfr derived GATA supernatant treated ES cells. There is F-actin accumulation in the epical epiblast domain and a lack of endoderm with strong ROCKII staining in H.