Fig. 6. Silencing SOD2 expression by siRNA increases autophagy and autophagic cell death induced by rotenone or TTFA in HeLa cells. (A) HeLa cells were transfected with scrambled or SOD2 siRNAs. The cells were lysed and western blotted for SOD2. The blot was stripped and re-probed for actin. (B-F) After HeLa cells were treated with rotenone (R, 50.0 µM) or TTFA (T, 0.5 mM) for 24 hours, (B) ROS generation, (C) AVO formation, (D) formation of GFP-LC3-labeled vacuoles (dots), (E) cell death and (F) apoptosis (formation of sub-G1 peaks) were determined as described in the Materials and Methods section. Error bars represent s.e. from three independent experiments. * Represents significant difference from control conditions (P<0.05). # Represents a lack of significant difference from control conditions.