Fig. 6.

TMZ-POH impairs lysosome and its fusion with autophagosome (a) A549 and SPC-A1 cells were treated with TMZ-POH with or without presence of Baf.A1, and incubated with LTR and detected using flow cytometry; (b, c) A549 and SPC-A1 cells were treated with TMZ, POH, TMZ + POH, TMZ-POH or DMSO respectively, and were analyzed by either western blot (b) or immunostaining (c) using a specific EEA 1 antibody. d Western blot analysis of CTSD in A549 and SPC-A1 cells treated with TMZ-POH or not; (e) Western blot demonstrated LAMP1 and LAMP2 protein level in A549 and SPC-A1 cells treated with TMZ-POH or not; (f) Relative location of autophagosome and lysosome in A549 cells treated with TMZ-POH or DMSO was observed by TEM. g A549 cells were either starved or treated with TMZ-POH, and stained with anti-LAMP1 and anti-LC3B antibodies and imaged by confocal microscopy. Statistical analysis of the colocalization coefficient of LAMP1 and LC3B.The results shown are means ±SD, *p < 0.05