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. 2021 Mar 29;10(4):534. doi: 10.3390/antiox10040534

Figure 2.

Figure 2

Effect of PE5 on autophagy induction and autophagic flux in lung cancer cells. (A,B) the morphological changes of the cells were detected using a microscope and the number of vacuoles per cells were calculated after H460 and H292 cells were treated with PE5. (C) H460 cells were treated with PE5 and stained with monodansylcadaverine (50 μmol/L) and visualized by fluorescence microscopy (Olympus IX51 with DP70). (D) H460 cells were treated with 50 μM PE5 for 24 h and observed by transmission electron microscopy. Arrowheads indicate the autophagosomes and the arrows show the vacuoles. (E) After getting treated with indicated amounts of PE5 for indicated time periods (6–48 h), LC3 proteins were measured by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. (F,G) Autophagy flux was determined in 50 μM PE5-treated cells in the presence of chloroquine (10 µM). The LC3 proteins were determined by western blot analysis. The blots were reprobed with GAPDH to confirm equal loading of the protein samples. The cells were treated 50 μM PE5 with or without chloroquine (10 µM) for 24–48 h. The level of LC3 expression was analyzed by immunofluorescence staining. Data represent the mean ± SD (n = 3) and (* p < 0.05, ** p < 0.01, compared with the untreated control) (# p < 0.05, compared with PE5-treated alone or different time).