Fig. 3

CDK13 interacts with E2F5 to cooperatively promote PCa cell proliferation. a and b, RT-qPCR and Western blot analysis detected the CDK13 expression in PC3 cells co-transfected with dCas9–VP64, MS2–p65–HSF1 and CDK13 promoter sgRNA (hereafter abbreviated as CDK13 sgRNA) or negative control (NC) sgRNA. *P < 0.05 vs NC sgRNA. c, Western blot analysis detected the E2F5, CDK13 and p21 expression in PC3 cells transfected with shE2F5, oeE2F5 or their respective control vector. d, Western blot analysis detected the proteins described as in (C) in PC3 and 22RV1 cells transfected with CDK13 sgRNA, shE2F5 or their respective control vector. e, Cells were prepared as in (D), cell viability was measured by MTS assay. *P < 0.05 vs. their respective controls. f, PC3 and 22RV1 cells were transfected with oeE2F5 and CDK13 sgRNA either alone or together, and colony formation assay was performed. Right panel shows the quantitative analysis of colony numbers from three independent experiments. *P < 0.05 vs. control constructs or CDK13 sgRNA and oeE2F5 alone. g, PC3 cells were transfected with CDK13 sgRNA or negative control (NC) sgRNA (left panel), as well as with siPol ll or siCtl (right panel). CoIP analysis detected the interaction between E2F5, CDK13 and Pol ll. h, Colony formation assay detected the proliferation of PC3 and 22RV1 cells co-transfected with the indicated constructs. Right panel shows the quantitative analysis of colony numbers from three independent experiments. *P < 0.05 vs. control constructs or siPol ll and CDK13 sgRNA alone