Fig. 2: Fluorizoline inhibits the removal of damaged mitochondria.

HeLa Parkin cells (CT) were treated with 20 μM pan-caspase inhibitor Q-VD-OPh (A, B, E, F) or not (C, D) while treated for 16 h with 1 μM oligomycin/1 μM antimycin (OA) or 10 μM CCCP, in the absence (UT) or presence of 10 μM fluorizoline (F) or 500 nM rocaglamide A (RocA). Cells were co-immunostained for DAPI (blue), TOM20 (green), and PHB2 (red) and co-localization was analyzed by confocal microscopy (A). The integrated density of TOM20 and PHB2 was quantified. Mean ± SEM (n = 3 independent experiments) (B). Protein levels from whole-cell lysates were analyzed by western blot and actin was used as a loading control (C). TOM20 and VDAC1 protein expression levels were quantified. Mean ± SEM (n = 3 independent experiments) (D). The genes for COX II and the amyloid precursor protein (APP) were amplified and measured by RT-qPCR as indicatives of mitochondrial and nuclear DNA, respectively. Mean ± SEM (n = 3 independent experiments) (E, F). These are representative images of three independent experiments (A, C). **p < 0.01 and ***p < 0.001 CT versus OA-treated cells (B); *p < 0.05, **p < 0.01 UT versus F or RocA-treated cells (D−F); ^##p < 0.01 CT versus CCCP and OA-treated cells (D).