Figure 2.

Pro-oxidant is able to push forward autophagic clearance of mitochondria. (a) HeLa cells stably expressing HA-Parkin were treated with DMSO, CCCP, antimycin A or H₂O₂ as depicted in (b). Cells were stained for hsp60 (red) and HA (purple). (c) Cells from (a) were counted as in Figures 1e and h. The error bars represent S.E.M. from three independent experiments. (d) HeLa cells expressing HA-Parkin were treated as indicated and immunoblotted for COX IV and GAPDH. Bottom: Average protein levels of COX IV relative to GAPDH from three independent experiments. (e) HeLa cells expressing HA-Parkin were transfected with scramble or FIP200 siRNA. Seventy-two hours post-transfection, cells were treated with DMSO or CCCP/antimycin A as in (a) for 24 h. Cells were stained for HA (green) and hsp60 (red). NS, not significant; *P<0.05; **P<0.01 (t-test)