Figure 2

Paclitaxel induced autophagy in UOK257 and ACHN-5968 cells. A. UOK257/UOK257-2 and ACHN-sc/ACHN 5968 cells were treated with 100 nM paclitaxel for 24 hours. In the absence or presence 100 nM bafilomycin A1 for four hours before harvest, elevated LC3-II levels were detected in FLCN-deficient UOK257 and ACHN-5968 cells by Western blot assay. B. In the absence or presence bafilomycin A1, LC3 protein levels were examined for cells treated with paclitaxel at various concentrations for 24 h, the highest LC3 level was observed at 100 nM in FLCN-deficient cells. C. FLCN-deficient cells were treated with 100 nM paclitaxel and harvested at different time intervals with or without bafilomycin A1 treatment. LC3-II expression peaked at 24 h treatment. D. Cells were treated with 100 nM paclitaxel and harvested with or without bafilomycin A1 treatment. In the absence of lysosomal inhibitor bafilomycin A1, decreased p62 was observed in paclitaxel-treated FLCN-deficient cells. E. Paclitaxel-induced autophagosomes in cells were observed using transmission electron microscopy. Autophagosome formation was found in FLCN-deficient UOK257 and ACHN-5968 cells. Arrows indicate autophagosome structures. Scale bars = 500 nm (*: p < 0.05. UOK257 vs UOK257-2; ACHN-sc vs ACHN 5968; n = 30). F. Cells were transfected with GFP-LC3 and analyzed under fluorescent microscopy for autophagosomes (*: p < 0.05, UOK257 vs UOK257-2; ACHN-sc vs ACHN 5968; n = 60). Scale bars = 15 μm.