CX-4945 induced the autophagy in EGFR-TKI-resistant PC-9 cells.

<p>A, Cells were treated with different concentrations of CX-4945 for 72 h, and the rate of inhibition was determined by MTT assay. B, Cells were treated with or without CX-4945 (5 µM) for 48 h and were then incubated for 24 h with a drug-free medium containing 10% FBS. Pictures showing the autophagic vacuole formation (AVOs) were taken at ×20 magnification. C and D, Cells were treated with CX-4945 for 48 h. Cell lysates were subjected to Western blot analysis. Quantitative detection of acidic vesicular organelles by acridine orange staining of cells was determined by FACS analysis. *p<0.01 and **p<0.001 compared with the control. E, Cells were transfected with a plasmid to express LC3-GFP. After 24 h transfection, cells were treated with CX-4945 (5 µM) for 24 h. Punctate pattern of LC3 localization analyzed by immunofluorescence microscopy. F and G, Cells were incubated with CX-4945 (5 µM), 17-DMAG (100 nM) or rapamycin (20 µM) for 48 h. Pictures were taken at ×20 magnification. The induction of LC3-I/II was shown by Western blot analysis.