Figure 1

Docetaxel induced autophagy in LAD cells. (A) SPC-A1 and H1299 cells were treated with either the indicated concentrations of docetaxel for 24 h or 10 μg/l docetaxel for varying periods. Whole cell lysates were subjected to western blot analysis for LC3, p62 and GAPDH expression (as a loading control). (B) Both parental cells (SPC-A1 and H1299) and docetaxel-resistant cells (SPC-A1/DTX and H1299/DTX) were transiently transfected with a GFP-LC3 construct. Twenty-four hours later, parental cells were exposed to docetaxel (10 μg/l) for an additional 24 h. GFP-LC3 dot formation was analyzed as described in Materials and Methods (mean ± S.D. of three independent experiments, *P < 0.05, **P < 0.01). Bar = 50 μm. (C) SPC-A1 and H1299 cells pretreated with or without bafilomycin A1 (20 nM, 2 h) were exposed to docetaxel (10 μg/l) for an additional 24 h. Whole cell lysates were analyzed by western blot. (D, E) Parental cells (SPC-A1 and H1299) and docetaxel-resistant cells (SPC-A1/DTX and H1299/DTX) were analyzed by (D) western blot analysis for LC3 and GAPDH expression and (E) transmission electron microscopy analysis, as described in Materials and methods. A magnified view of the electron photomicrograph showed the characteristic autophagosomes. N: nucleus. The results were obtained from three independent experiments.