Fig. 1

TMZ-POH induces autophagosome formation. a, b Cells were treated with 100 μM TMZ, POH, TMZ + POH, TMZ-POH or DMSO respectively for 48 h. a Western blot analysis demonstrated LC3B and ACTB expression in above drug-treated A549, SPC-A1, H460 and H520 cells; (b) The above drug-treated cells were inspected under confocal laser microscopy to detect LC3B puncta by immunofluorescence. LC3B puncta number per cell was quantified using the Fiji Image J program; (c) Autophagic vacuoles in A549 cells treated with 100 μM TMZ-POH or DMSO were observed by transmission electron microscopy (TEM). The arrow indicates autophagic vacuoles. Number of autophagic vacuoles were calculated using Fiji Image J software. d SPC-A1 cells treated with 100 μM TMZ-POH or DMSO were inspected under confocal laser microscopy to detect LC3B puncta by immunofluorescence in the presence or absence of Baf.A1. The results shown are means ±SD, **p < 0.005, ***p < 0.001, NS = no significance