CHD4 and MBD3 regulate the acetylation and protein levels of p53.
A, qRT-PCR analysis. Each mRNA level was normalized to the β-actin level, and the value of control shRNA-expressing cells at day 4 was set to 1. The data are shown as the means ± S.D. (n = 3 independent experiments). **, p < 0.01. The p values were calculated using Student's unpaired two-tailed t tests compared with control cells on the same day. B, immunoblotting analysis for acetyl-p53 (Lys-379), p53, p300, CHD4, HDAC1, Myc, and α-tubulin (as a loading control) in lysates from HEK293T cells transfected with the indicated combination of expression vectors (left). The relative acetyl-p53 (Lys-379) signal intensities are shown (right). The data are shown as the means ± S.D. (n = 3 independent experiments). *, p < 0.05, and **, p < 0.01. N.S., not significant. The p values were calculated using Student's unpaired two-tailed t tests. C, immunoblotting analysis for p53, acetyl-p53 (Lys-379), and α-tubulin (as a loading control) in control shRNA- and Chd4 shRNA-expressing ESCs at day 0 and day 4 of SFEBq culture. D, immunoblotting analysis for p53, acetyl-p53 (Lys-379), and α-tubulin (as a loading control) in control shRNA- and Mbd3 shRNA-expressing ESCs at day 0 and day 4 of SFEBq culture.