Treatment with JQ1 reduces BRD4 and Pol II occupancy on the promoters of c-MYC and BCL2, and depletes the mRNA expression of c-MYC and BCL2 in human MCL cells. (A-B) MO2058 cells were treated with the indicated concentrations of JQ1 for 16 hours. Following this, ChIP was conducted with a BRD4-specific antibody (A) or RNAP2 antibody (B). The ChIP DNA was subjected to real-time qPCR with primers against the enhancer and promoter of c-MYC and the promoter of BCL2. The fold-change was calculated using the cycle threshold (Ct) value of the ChIP DNA compared with the Ct value of the input DNA. (C) MO2058 cells were treated with 1000 nM of JQ1 for 8 hours. Total RNA was extracted and used for gene expression analyses. A heat map of TNFAIP3, MYC, CDK4, BCL2, CDK6, and HEXIM1 is shown. (D) MO2058 cells were treated with the indicated concentrations of JQ1 for 16 hours. At the end of treatment, RNA was isolated and reverse transcribed. The resulting cDNA was used for real-time qPCR analysis of c-MYC, BCL-2, and CDK6. The relative mRNA expression was normalized to GAPDH and compared with the untreated cells. (E) Representative immunoblots of MO2058 cells treated with the indicated concentrations of JQ1 for 24 hours. Immunoblot analyses were conducted for the expression levels of c-MYC, MCL1, CDK4, CDK6, HEXIM1, p21, p27, BIM, and β-actin in the cell lysates.