Fig. 3
Prostate cancer cells with 17p deletion are highly sensitive to RBX1 depletion. a, b Effect of RBX1 knockdown on the proliferation of the parental and isogenic 17ploss DU145 cells, determined by direct competition assay. Cells expressing RFP and control nonspecific shRNA (shNT) or shRBX1 were sorted and mixed with control RFP-negative cells (1:1) and the RFP-positive cells were quantified at passages 2, 4, and 6 (a). Representative cell survival measured by staining with crystal violet was shown in b. c–e Cell growth curves, based on crystal violet staining, of human prostate cancer cell lines expressing Dox-inducible shNT or RBX1-specific shRNA (shRBX1 #1, shRBX1 #2) (c). RBX1 knockdown efficiency and representative image were shown in d and e, respectively. f Fraction of apoptotic cells in the 17pneutral (22Rv1 and DU145) and 17ploss (PC3 and VCaP) cells expressing Dox-induced RBX1 shRNA at 4 days post Dox treatment. g, h Cell survival measured by crystal violet staining (g) and protein expression levels (h) of RBX1 in PC3 and VCaP cells expressing shRBX1, control or ectopic RBX1. Data are representative of three independent experiments and analyze by unpaired two-tailed t-test. Error bars denote SD. **, p < 0.01; ***, p < 0.001
