Fig. 2

miR-519a enhanced TMZ-induced autophagy in GBM cells. The expression levels of LC3-II in U87-MG/TMZ and U87-MG cells were evaluated by immunofluorescence assays (a) and Western blotting (b). Red indicates LC3B and blue indicates nuclei. c Both U87-MG/TMZ and U87-MG cells were transfected with GFP-LC3 construct expressing either miR-519a or anti-miR-519a, followed by treatment with 400 μM TMZ. The numbers of GFP-LC3 puncta were quantified using confocal laser scanning microscopy. d Both U87-MG/TMZ and U87-MG cells were transfected with either miR-519a or anti-miR-519a for 24 h, followed by treatment with 400 μM TMZ. Cell samples were prepared for transmission electron microscopy analysis. The arrows indicate autophagic vacuoles. e U87-MG/TMZ cells transfected with miR-519a and parental U87-MG cells transfected with anti-miR-519a were exposed to 400 μM TMZ at different time points (0–24 h). Whole cell lysates were analyzed by Western blotting. f U87-MG/TMZ cells transfected with or without miR-519a and treated with or without 400 μM TMZ after incubation with 20 nM bafilomycin A1 for 2 h. U87-MG cells transfected with or without anti-miR-519a were treated with 20 nM bafilomycin A1 for 2 h. Cell lysates were analyzed by Western blotting. *p < 0.05, **p < 0.01