Pcgf1 had a positive role in maintaining the pluripotency of P19 cells. (A) qPCR showed that the expression of Pcgf1 was increased in P19 cells induced by retinoic acid, especially 24 h later. (B,C) The expression of neural markers Pou3f1 and Zfp521 were increased, while the pluripotent markers Oct4 and Nanog were decreased. (D) Construction of stable P19 cell line with Pcgf1 knockdown by lentivirus. (E) P19 cells clustered earlier than the control group after Pcgf1 knocked down at 24 and 48 h induced by RA. Scale bar, 100 μm. (F) The expression levels of the neural markers Pou3f1 and Zfp521, and the pluripotent markers Hes1 and Nanog, were detected by qPCR after Pcgf1 knock down at 2 and 48 h induced by RA. (G) The expression levels of the neural marker Pax6 and the pluripotent marker Oct4 were detected by Western blot after Pcgf1 knock down at 0, 24, and 48 h induced by RA. (H) The expression levels of the neural marker Pax6 and the pluripotent marker Nanog were detected by qPCR after overexpressed Pcgf1 at 24 and 48 h was induced by RA. (I) The Western blot results showed that pluripotent marker Oct4 was consistently expressed after overexpressed Pcgf1 at 0, 24, and 48 h was induced by RA. Data represent the mean of at least three independent experiments ± SEM. The expression of β-actin gene represented internal controls. *P < 0.05 vs. control, **P < 0.01 vs. control, ***P < 0.001 vs. control.
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