Research Paper Volume 12, Issue 23 pp 24208—24218

Figure 3. The anti-carcinoma activity of SRT2183 is dependent on autophagy. (A) OVCAR-3 and A2780 cells were pre-treated with chloroquine (CQ) for 6 h, then treated with vehicle or 1 μM SRT2183. The expression of LC3, p62/SQSTM1, Caspase3, PARP and Bcl-2 was detected by western blot. (B) OVCAR-3 and A2780 cells were treated by the same procedure described in (A) for 24 and 48 h, the viability of OVCAR-3 and A2780 cells were detected by CCK8 assay. (C) The expression of autophagy related 5 (ATG5) was knocked down by shRNA-lentivirus (shATG5-1 and shATG5-2) and the protein was determined by western blot. (D) OVCAR-3 and A2780 cells were treated by vehicle, shATG5-1, 1 μM SRT218 or the combination of shATG5-1 and SRT218 for 24 and 48 h. The viability of the cells was evaluated by CCK8 assay. (E) The expression of autophagy related 5 (ATG5) was detected by western blot and (F) The cell viability was determined after the cells were treated by 1 μM SRT2183 for 24 and 48 h. (G, H) OVCAR-3 and A2780 cells were pre-treated with 0.1 μM Rapamycin (Rapa) or 0.05 μM Trion 1 for 6 h, then the cells were treated with vehicle or 1 μM SRT2183 for 24 and 48 h. The cell viability was detected by CCK8 assay. N=3 for (A) and (C); N=5 for (B), N=6 for (D), (E) and (F). * indicates P < 0.05; ** indicates P <0.01; *** indicates P < 0.001.