Fig. 3

PPFIA4 promotes PCa cell growth in vitro and in vivo. A-D. Cell proliferation was assessed by cell counts and colony formation assays in C4-2B cells transfected with siNC or PPFIA4 siRNA (siPPFIA4) (A-B) or in LNCaP cells transfected with empty vector or PPFIA4 overexpression plasmid with or without androgen deprivation (C-D). Cell number was counted at the indicated time points and all the numbers were normalized to day 0 (A, C). Data shown are presented as means ± SEM of triplicate wells and are representative of at least three replicate experiments. Representative images of colony formation assays are shown in the left panel and quantitative analysis is shown in the right panel (B, D). **p < 0.01, ***p < 0.001, ****p < 0.0001 based on the Student’s t-test. d, days. FBS, fatal bovine serum. CSS, charcoal-stripped serum. E-H. EdU assays (E-F) and cell apoptosis assays (G-H) were performed in indicated PCa cells with PPFIA4 knockdown or overexpression. Representative images are shown in the left panel and quantitative analysis is shown in the right panel. * p < 0.05. **p < 0.01, ***p < 0.001, ****p < 0.0001. FBS, fatal bovine serum. CSS, charcoal-stripped serum. I-L. The tumor volume was measured in nude mice with castration as the indicated treatment. LNCaP cells with stable overexpression of PPFIA4 or C4-2B cells with stable knockdown of PPFIA4, as well as their parental controls, were subcutaneously injected into nude mice (n = 5/group). The tumor volume was measured twice every week after nude mice were castrated (I, K). Tumors were isolated from mice, photographed, and weighed at the endpoint (J, L). ***p < 0.001