Fig. 5: ACLY promotes P300 expression to accelerate H3K27 acetylation of Nanog promoter in SF CRC cells.

A The expression levels of H3K27Ac were analyzed by Western blotting in normal cells, in SF CRC cells and in SF CRC cells treated with ETO. B The interaction of H3K27Ac and Nanog promoter in SF CRC cells was assessed by CHIP-PCR. IgG was used as negative control. Scheme of the different positions of ChIP-PCR primers was also shown (P1: −1844/−823 bp, P2: −1043/−1024 bp, P3: −791/−772 bp, P4: −259/−236 bp). C The expression levels of histone acetyltransferases (P300, KAT2A, KAT3A) and histone deacetylases (HDAC1, HDAC2) were examined by PCR in normal HT29, in SF HT29 cells and in SF HT29 cells treated with ETO. D The expression level of P300 was analyzed by Western blotting in normal CRC cells, in serum-deprived CRC cells and in serum-deprived CRC cells treated with ETO. E, F P300 inhibition downregulated the expression of Nanog and H3K27Ac. The serum-deprived HT29 and HCT116 cells were treated with C646 for 24 h. Then, the cells were tested for Nanog expression by PCR (E) and Nanog and H3K27Ac expression by Western blotting (F). C646: P300 inhibitor. Data are shown as the means ± s.e.m., n = 3; ns: no statistical significance, **P < 0.01, ***P < 0.001. Data shown represented three independent experiments.