Fig. 3.

RNA interference-mediated autophagy blockade reduces the viability of ascites-derived ovarian cancer cells. The iOvCa147-E2 cell line was transfected with non-targeting siRNAs (siNT) or siRNA pools targeting BECN1, ATG5 and ATG7. Transfection was repeated 72h later, and following overnight incubation, cells were seeded for subsequent experiments. (A) At 10 days after initial transfection, iOvCa147-E2 cells were treated for 24h with either DMSO or Akti-1/2 (3.5 μM). Lysates were obtained and immunoblots performed for indicated proteins. Blot represents four repeated experiments. (B) At 5 days post-transfection, cells were treated with DMSO vehicle control or Akti-1/2 (3.5 μM) in quadruplicate and viability assessed by alamarBlue assay at indicated time points. Data are pooled from three experiments and represented as mean ± SEM.