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. 2021 Jan 23;10(4):1405–1417. doi: 10.1002/cam4.3734

FIGURE 1.

FIGURE 1

EGFR‐TKI treatment activates autophagy in PC9 and PC9/GR cells. (A) Western blot analysis of lysates derived from PC9 and PC9/GR cells treated for 6 h with increasing concentration of gefitinib. (B) Analysis of LC 3 aggregation in PC9 and PC9/GR cells by fluorescence microscopy. The LC3 puncta were determined after treatment of 0, 0.1, 1, and 10 μM gefitinib. At a high dose of gefitinib, most of PC9 cells died (Scale bar, 200 µm). (C) Immunofluorescent staining of PC9 and PC9/GR cells transfected with mRFP‐GFP tandem fluorescent‐tagged LC3 construct. Chloroquine blocked the fusion of autophagosome and lysosome, causing GFP accumulation, however, gefitinib caused a significant decrease of GFP in both cell lines, indicating the activation of autophagic flux. (D)TEM images of PC9 and PC9/GR cells showed that gefitinib treatment increased cytoplasmic organelles with a double lumen in both cell lines, suggesting activation of autophagy. Representative images are shown, and the arrows indicate autophagosomes (scale bar, 1 µm)