Fig. 4

PD0325901 blocked formation of the primitive streak by preventing β-catenin accumulation in the nucleus. a Immunostaining showed that PD0325901 could not prevent the translocation of SMAD2 into nucleus in EpiSCs induced by Activin A. Bar, 100 μm. b Inhibition of phospho-ERK1/2 by PD0325901 did not affect phospho-SMAD2 and SMAD4 detected by western blot analysis. c Locations of SMAD2/3/4 were not changed in EpiSCs after PD0325901 treatment for 24 hours. C cytoplasm, N nucleus. d Western blot analysis showed that PD0325901 inhibited translocation of β-catenin into the nucleus. XAV939 (XAV, 2 μM) was used as a positive control that promoted the retention of β-catenin in cytoplasm. e PD0325901 increased expression of E-cadherin protein upon N2B27, CHIR99021, and Activin A treatment. SMAD SMAD family member, DMSO dimethyl sulfoxide, PD PD0325901, ERK extracellular signal-regulated protein kinase, SOX2 sex determining region Y-box 2, CHIR CHIR99021, OCT4 octamer-binding transcription factor 4, T brachyury