Figure 7.

CHD4 regulates neural differentiation through p53 down-regulation. A, immunoblotting analysis for p53 and α-tubulin (as a loading control) in Chd4 shRNA #2- and/or p53 shRNA-expressing cells at day 0 and day 4 of SFEBq culture. B, effect of Chd4 and/or p53 shRNAs on the number of ESCs. One day before viral infection (day −3), 2.5 × 10⁵ cells were plated, and the cells were counted at day 0. C, qRT-PCR analysis. Each mRNA level was normalized to the β-actin level, and the value of control shRNA-expressing cells at the 0 was set to 1. D, SFEBq-cultured cells at day 4 were transferred to dishes for further neural differentiation. Three days later (day 7), cells were stained for TUJ1 (upper). The scale bars represent 100 μm. The percentages of TUJ1-positive cells are shown (lower). B–D, data are shown as the means ± S.D. (n = 4 independent experiments for C, and n = 3 independent experiments for B and D). *, p < 0.05, and **, p < 0.01. N.S., not significant. The p values were calculated using Student's unpaired two-tailed t tests compared with control cells on the same day. In addition, the p values were calculated using Student's unpaired two-tailed t tests between Chd4 shRNA-expressing cells and Chd4 and p53 shRNA-expressing cells on the same day.