Notch3 Interactome Analysis Identified WWP2 as a Negative Regulator of Notch3 Signaling in Ovarian Cancer (original) (raw)
Figure 6
WWP2 regulates Notch3 signaling activity in cancer cells.
(A) OVCAR3 or MCF7 cancer cells were transfected with control plasmid pLPC, WWP2, catalytically-inactive WWP2 (WWP2-CA), N3-ICD, or WWP2+N3-ICD. Relative cell numbers were measured at different time points using the SYBE Green-based assay. Data are expressed as means ± SD and Student's _t_-test was performed to compare the relative cell number between WWP2+N3-ICD and pLPC (* p<0.05; ** p<0.01). (B) Different groups of cells were transfected with the pJH23A (4×wtCBF1Luc) luciferase reporter construct together with pLPC, WWP2, or the catalytically-inactive WWP2 plasmid (WWP2-CA). Each experiment was performed in triplicate. Results are shown as percentage of luciferase activity compared to pLPC transfected controls. Data are expressed as means ± SD. P values were calculated by comparison between either WWP2 or WWP2-CA and the control plasmid pLPC (** p<0.01; *** p<0.001). (C) Two untransformed epithelial cell lines were transfected with WWP2 siRNA or scrambled siRNA (siSCR). Notch signaling was measured by co-transfecting the cells with a luciferase reporter plasmid, pJH23A (4×wtRBPJLuc) and the data were normalized to the luciferase activity measured by co-transfection with a control plasmid, pGL3. Data are expressed as means ± SD and statistical significance was assessed by two-tailed Student's _t_-test.