Fig. 4

Effect of NRP1 silencing on Olaparib resistance. SKOV3 cells were transfected with NRP1-specific siRNA (siNRP) or nonspecific control siRNA (siNC). a, b) NRP1 expression was assessed at 48, 72 or 144 h from transfection (a), and after Olaparib treatment at 144 h (b) by Western blot analysis. Tubulin expression was used as internal control. The images are representative of at least three independent experiments. c, d) The effect of Olaparib on cell viability and clonogenicity of silenced SKOV3 cells was determined by MTT assay and colony formation assay, respectively. Mean values obtained from two independent experiments, each performed in triplicate, are reported in graph. Error bars represent standard deviations. **, p < 0.005, ***, p < 0.0005 vs. siNC-transfected cells or vs. control (DMSO). e) The expression of the apoptosis related proteins phospho-AKT (Ser473), AKT, cleaved caspase 3 and cleaved PARP1 in silenced SKOV3 cells treated or not with Olaparib was assessed by Western blot analysis. Tubulin expression was used as internal control. The images are representative of at least two independent experiments