(A and B) Western blot and BrdU incorporation analysis on (A) PC3 and (B) LNCaP cells (*P < 0.001; n = 3, analyzed by Mann-Whitney test). (C and D) Representative orthotopic tumor images and the weights of prostate following injection with (C) DU145 and (D) LNCaP cells as indicated. (*P < 0.01, number of mice = 5, analyzed by Dunnett’s multiple comparison test). Scale bars: 0.5 cm. Box and whisker plots show median (lines within boxes), interquartile range (bounds of boxes), and upper and lower range (whiskers). (E) SPRY2 KD DU145 were analyzed for cell-cycle profiles (top panel) after thymidine block release, and percentage of G1 cells are in bar chart (bottom panel; *P < 0.01; n = 3, analyzed by Mann-Whitney test). (F) SPRY2 KD DU145 transfected with PTEN siRNA were analyzed by Western blot and the G1 cells quantified. (*P < 0.01; n = 3, analyzed by Mann-Whitney test). (G) MEFs as indicated were quantified for cells in G1 (*P < 0.001; **P < 0.01; n = 3, analyzed by Mann-Whitney test). All Western blots were quantified using ImageJ, and the values represent relative immunoreactivity of each protein normalized to respective loading control. Data (A, B, F, and G) presented as mean ± SEM.