Figure 4.
BET-BRD inhibition with JQ1 potently suppresses c-Myc gene expression in Pkd1 mutant renal epithelial cells. (A) CHIP assay was performed with anti-Brd4 antibody or anti-histone 3 antibody in Pkd1 mutant PN24 cells treated with DMSO or JQ1 (1 µm) for 4 h, respectively. The precipitated chromatin DNA was analyzed by quantitative PCR with primers that amplified from −613 and −374 bp upstream of the c-Myc ATG start codon. Enrichment relative to no antibody control was shown. n = 3, *P < 0.05. (B and C) qRT-PCR analysis of the expression of Brd4 and c-Myc mRNA in postnatal Pkd1 mutant PN24 cells (B) and Pkd1 null MEK cells (C) treated with 1 µm JQ1 for 24 h. n = 3, **P < 0.01. (D) Western blot analysis of the expression of c-Myc in Pkd1 mutant PN24 cells treated with 1 µm JQ1 at indicated time points. (E and F) Western blot analysis of the expression of c-Myc in Pkd1 mutant PN24 cells (E) and Pkd1 null MEK cells (F) treated with JQ1 with indicated concentrations.