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. 2015 Sep 5;6(32):33397–33409. doi: 10.18632/oncotarget.5179

Figure 6. JQ1 directly suppresses Bcl-xL and Rad51 in Gnaq/11 mutant cells.

Figure 6

The effect of JQ1 on the mRNA of Bcl-xL and Rad51 was confirmed by qPCR in UM cell lines. Total RNA was extracted from cells with different mutational status after 24 h of treatment with 500 nM JQ1, and qPCR was performed using gene-specific primers for Bcl-xL A. and Rad51 B. Values were normalized with GAPDH as housekeeping gene using the ΔΔCT method. Values are relative to mRNA levels of 92.1 untreated cells set at 1. Each experiment was performed two or three times in triplicates. Bars, ± sd. *, **P < 0.01; #P < 0.05, comparing treatment versus DMSO. BRD4 ChIP assay for the Bcl-xL promoter C. and Rad51 promoter D. presented as percent of input, before and after treatment. Bars are representative of two independent experiments ± sd. *P < 0.001; **P < 0.01; #, P < 0.05. E. Downregulation of BRD4 regulates Bcl-xL and Rad51 expression in the Gnaq-mutant cells. The indicated cell lines were transfected with a non-specific siRNA (−) or BRD4 siRNA (+), and analyzed by immunoblotting with the indicated antibodies.