Fig. 6
Silencing of ZNF322A induces autophagosome formation in lung cancer cells. a-b Protein expression of mTOR and p-mTORS2448 was determined and normalized by immunoblotting. c-d Induction of autophagy related proteins including LC3B-I, LC3B-II, and SQSTM1 were analyzed and quantified by immunoblotting. Earle’s balanced salts solution (EBSS) was used as positive control to induce autophagy. e-f Electron microscopy of A549 lung cancer cells after 24 or 48 h of siControl or ZNF322A siRNA transfection was analyzed. Double- and multi-membrane autophagosomes were clearly observed in ZNF322A-silenced cells. Final stages of autophagic cell death were accompanied by ballooning of the perinuclear space and disappearance of cellular organelles such as autophagosomes, autolysosomes, and ER. Arrows: diverse autophagosomes, N: nuclear. β-actin is the internal control to normalize protein expression. The bars represent densitometric analysis of three biological replicates and the data are shown as mean ± SD. * p < 0.05; ** p < 0.01