Fig. 2.

SPOP regulates endogenous CYCLIN E1 specific in prostate and bladder cancer cell lines. a, b The indicated cell lines were transfected with control or SPOP-specific siRNAs. After 72 h, cells were harvested for WB. c After treatment with DMSO or MG132 (10 mM) for 12 h, DU145 lysates were prepared for co-IP with CYCLIN E1 antibody. d DU145 and PC3 cells were transfected with HA- or Myc-SPOP plasmid. After 24 h, cells were treated with DMSO or MG132 (10 mM) for 12 h and endogenous CYCLIN E1 level was assayed with WB. e Flag-SPOP (green) was expressed in PC3 cells and endogenous CYCLIN E1 (red) was assayed by immunofluorescent staining. f SPOP was knocked down in DU145 cells, and 48 h later cells were treated with 100 μg/ml cycloheximide (CHX), and harvested at various time points for WB. CYCLIN E1 protein abundance was quantified by ImageJ and plotted as indicated. *means p-value < 0.05