Figure 8

(A) Representative TEM images of A549 cells treated with 4a and 4b at the indicated concentrations for 24 h. Scale bars: 5 μm. (B) Representative images of A549 cells expressing GFP-LC3 treated with 4a (4 μM), 4b (4 μM) and rapamycin (1 μM) for 24 h. Scale bar: 5 μm. (C) Western blot analysis of LC3 proteins extracted from A549 cells. Cells were treated with 4a (4 μM) and 4b (4 μM) for 24 h. (D) Confocal microscopic analysis of A549 cells stained with AO after treatment with 4a (4 μM) and 4b (4 μM) for 24 h. AO was excited at 488 nm and monitored simultaneously at 500‒550 nm (Green) and 670‒710 nm (Red). Scale bars: 10 μm. (E) Mean red fluorescence intensity determined by flow cytometry in A549 cells stained with AO after treatment with 4a or 4b at the indicated concentrations for 24 h. AO was excited at 488 nm and emission was collected at 670‒710 nm. (F) The impact of 3-MA on anti-proliferative activity of cisplatin, 4a and 4b. Cells were pre-treated with 3-MA for 1 h and then incubated with cisplatin, 4a or 4b at indicated concentrations for 24 h. **p < 0.02.