SR9009 inhibited autophagy in SCLC cells. (A) After stained with Cyto-ID Green dye, the formation of autophagic vesicles in H69AR and H446DDP cells treated with or without SR9009 for 48 h was detected by laser confocal microscopy. Quantification of autophagic vesicles (Cyto-ID) in cells were measured by the ImageJ software (n = 3, means ± S.D.). Confocal micrographs were taken at × 40. (B) H69AR and H446DDP cells were incubated with or without SR9009 for 48 h and TEM was employed to detect the autophagosomes, and the magnified view of the electron photomicrograph exhibited autophagosomes. Arrows, autophagosomes. A typical result from 3 independent experiments is presented. (C and D) H69AR and H446DDP cells were incubated with different concentrations of SR9009 for 48 h, then autophagy associate protein LC3-I/II and P62 was detected by western blot analysis. (E and F) H69 and H446 cells were incubated with different concentrations of SR9009 for 48 h, then LC3-I/II and P62 was detected by western blot analysis. (G and H) Representative IHC staining of autophagy associate protein P62 in subcutaneous xenografts of SR9009-treated group or vehicle group.