Figure 3.

Blocking PLK4 or STIL Promotes Stem Cell Differentiation

(A) Phase-contrast images of hESCs or hiPSCs after 8 or 4 days of treatment, respectively. Arrows point to observed morphological changes. Scale bars, 50 μm.

(B) Analyses of mRNA levels of T, GATA6, and PAX6 in hESCs and hESCs #2 after 4 days of treatment. Data are presented as relative fold change over control.

(C–E) Western blot analyses of hESCs and hiPSCs after indicated time of treatment, with α-tubulin as a loading control. (C) Analyses of effects on pluripotency and differentiation by the indicated antibodies. (D) Analyses of effects of treatment (2 days) on protein turnover of p53 after indicated time (hours) of inhibition of translation by cycloheximide (CHX). (E) Analyses of the effect of temporal mitotic arrest by 6 hr of nocodazole treatment. Left panel shows scheme of the experiment. Controls (asynchronous cells) and treated samples (Noco+shake off, Noco-leftover) were probed for protein levels of p53, brachyury, GATA-6, and PAX-6 2 days after nocodazole washout. Noco-leftover condition represents non-mitotic nocodazole-treated cells.

Data are presented as mean ± SEM (^∗p < 0.05, ^∗∗∗p < 0.001). See also Figure S3.