Effect of Pcgf1 on neural stem cells (NSCs) in neural tubes. (A) The expression levels of Sox2, Otx2, Ngn1, and Sox3 were detected by whole-mount in situ hybridization at self-renewal phase of NSCs (16 and 24 hpf). Scale bar: 200 μm. (B) The expression of Sox2, Otx2, Ngn1, and Sox3 was analyzed by qPCR, and the expression of β-actin gene represented internal controls. Data represent the mean of at least three independent experiments ± SEM, *P < 0.05 vs. control MO. (C) Acridine orange staining was used to detect apoptosis at 16 and 24 hpf; BrdU labeling of control MO and Pcgf1 MO-injected embryos at 16 and 24 hpf. The expression of PCNA was detected by whole-mount in situ hybridization. Data represent the mean of at least three independent experiments ± SEM, **P < 0.01 vs. control MO. (D) The expression of p21 and p57 were analyzed by qPCR, and the expression of β-actin gene represented internal controls. Data represent the mean of at least three independent experiments ± SEM. *P < 0.05 vs. control MO.
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