Fig. 5
EHMT2 directly binds to the APC promoter to suppress its expression. A EHMT2 binding peak on the promoter of APC. Public ChIP-seq data using EHMH2 antibody in HepG2 cells are from the ENCODE database. The red rectangle indicates the binding signal peaks. B ChIP-PCR showed that EHMT2 directly binds to the APC promoter. 1.0 × 107 Hep3B or Huhl cells were subjected to ChIP using the EHMT2 antibody (RLP30 serves as negative control). n = 4 (mean ± SD); n.s, no significance and ***p < 0.001 by student's t-test. C ChIP-PCR showed that the H3K9me2 level of APC promoter. Hep3B or Huhl in 2B was treated with UNC0642 at a dose of 5.0 μM for 24 h, and then 1.0 × 107 cells were subjected to ChIP using the H3K9me2 antibody (IgG control is GFP antibody). n = 4 (mean ± SD); n.s, no significance and ***p < 0.001 by student's t-test. D ChIP-PCR showed that PolII directly binds the APC promoter in Hep3B or Huhl cells. Cells and treatment were the same as 5C. 1.0 × 107 cells were subjected to ChIP using the PolII antibody (IgG control is GFP antibody). n = 4 (mean ± SD); n.s, no significance, *p < 0.05 and **p < 0.01 by student's t-test. E RT-qPCR showed APC expression in Hep3B and Huh1 cells. Cells and treatment were the same as 5C. n = 6 (mean ± SD); ***p < 0.001 by student's t-test. F The APC protein level in cells was determined by Western blot. Hep3B and Huh1 were treated with UNC0642 at the indicated concentrations. n = 6 (mean ± SD); ***p < 0.001 by one-way ANOVA