Figure 7.

Autophagy flux detection upon treating with Chloroquine (CQ) or Rapamycin (Rap) in wild type and p62 silencing A549 cells. (A) Double immunofluorescent staining of p62 and LC3 protein in A549 cells treated with CQ (20μM/L for 48h). Punctate GFP-LC3 emits green fluorescence, and punctuate mCherry-p62 emits red fluorescence. In merged panel p62 protein only partly colocalized with LC3 protein. (B) Changes of p62 and LC3 protein was detected upon Rap or CQ in different A549 cells. Significant switch of LC3 I to LC3 II was detected in different A549 cells treated with CQ or Rap, which demonstrates that autophagy flux is activated. Meanwhile, increased p62 expression is induced instead of degradation as autophagy substrate. In p62 silencing A549 cells, Significant switch of LC3 I to LC3 II was detected upon treating with CQ but not with Rap. Ctrl, untreated A549 cells; siR-p62, small interfering p62 RNA transfected cells; siR-NC, siR-p62 scrambled control; CQ, Chloroquine; Rap, Rapamycin. ^*P < 0.05, ^**P < 0.01.