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. 2020 Jul 13;219(9):e201910041. doi: 10.1083/jcb.201910041

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© 2020 Qi et al.

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Figure 6. — PTEN promotes epiblast differentiation and polarization via down-regulation of the WRC. (a and b) 5-d wild-type GFP and Pten⁻^−/− EBs stably transfected with Nckap1 shRNA or the scrambled control (SC) were analyzed by immunoblotting. Blots were quantified by densitometry, and the ratio of Nanog, Oct4, and keratin 8 to actin was plotted. Mean ± SD. *, P < 0.05 versus WT GFP and Pten^−/− Nckap1 knockdown (KD). (c) 5-day EBs were coimmunostained for MPDZ and perlecan. (d) EBs with polarized epiblast epithelium were counted and plotted as a percentage of total EBs examined. n = 1,090–1,166 for each group. Mean ± SD. (e and f) 5-d normal EBs stably transfected either with pCXN2-WAVE2 or only the vector were subjected to immunoblot analysis. Blots were quantified and the ratio of Nanog, Oct4, and keratin 8 to actin was plotted. Mean ± SD. *, P < 0.05 versus the vector control. (g) 5-d EBs were immunostained for MPDZ and perlecan. (h) Formation of the polarized epiblast epithelium in 5-d EBs were quantified by phase-contrast microscopy and plotted as a percentage of total EBs examined. n = 724–750 for each group. Mean ± SD. *, P < 0.01 versus control.