Figure 4. Atg5 and Atg7 play roles in induction of autophagy caused by downregulation of αSNAP. (A and B) SK-CO15 cells were subjected to sequential transfections with one of the following siRNA pairs: control-control, control-Atg5, control-Atg7, control-αSNAP, Atg5-αSNAP or Atg7-αSNAP. Expression of LC3, αSNAP, Atg5 and Atg7 was determined by immunoblotting 48 h after the second transfection. #p < 0.01 compared with control-αSNAP siRNA-transfected cells. (C and D) HeLa-GFP-LC3 cells were sequentially transfected with control-control, control-Atg5, control-αSNAP and Atg5-αSNAP siRNA combinations and formation of autophagosomes was analyzed by fluorescence spectroscopy at 72 h after the second transfection. *p < 0.001 compared with control-αSNAP siRNA-transfected cells. Scale bar, 20 µm.