Fig. 3

HMGA2 knockdown-induced inhibition of autophagy indirectly promotes NF1 MPNST cell apoptosis (a) Cells transfected with shHMGA2 exhibited a punctate pattern of LC3-II fluorescence, with reduced LC3-II expression compared with that in autophagosomes. b Representative transmission electron microscopy images depicting the ultrastructures present during autophagy in sNF96.2 and ST8814 cells transfected with shHMGA2 or shScr for 48 h. The images show autophagic vacuoles (arrows) in control cells. No or few autophagic vacuoles were observed in cells transfected with shHMGA2 or treated with 3MA. c WB analysis was used to evaluate the expression levels of LC3-II, p62 ATG7, ATG12 and Beclin1. d EdU assay revealed that the treatment of cells with rapamycin increased the number of viable HMGA2 knockdown cells. e TUNEL positivity of HMGA2 knockdown cells was markedly decreased in the presence of rapamycin. Scale bar = 50 μm. f Treatment with rapamycin markedly increased LC3-II, ATG7, ATG12, Beclin1 and BCL2 levels and decreased Bax and p62 levels. Data are presented as the mean ± SD. (n = 3). *P < 0.05 by Student’s t-tests