Figure 6.

Changes in detection of p62 and LC3 II/I protein upon induction or inhibition of autophagy. (A) In untreated wild type A549 cells, punctate GFP-LC3 (white arrow) was rare (left image) but increased significantly after exposure to rapamycin (10 μM/L) for 24 h (right image) by GFP-LC3 immunofluorescence microscopy. (B) Similar to wild type SPC-A-1 cells, punctate GFP-LC3 increased slightly after treatment with rapamycin. (C) Punctate GFP-LC3 positive cells were increased after rapamycin exposure both in A549 and SPC-A-1 cells. (D) p62 and LC3 II/I protein expression upon different concentrations of exposure to rapamycin for 24h. (E) Densitometric analysis of protein bands showing LC3 II/I protein expression increased from 5μM/L to 10μM/L and decreased from 50 μM/L to 100 μM/L, but p62 protein expression changes were not obvious. (F) Changes in detection of p62 and LC3 II/I protein upon autophagic gene 7 knockout (Atg7-/-). (G) Densitometric analysis of protein bands showing LC3 II/I protein expression almost disappeared after Atg7-/- knockout, but p62 protein expression decreased obviously both after autophagy induction by Rapamycin and autophagy inhibition by Atg7-/- knockout. Scale bar was 20 μm. wt, wild type A549 or SPC-A-1 cells; Rap, Rapamycin. ^*P < 0.05, ^**P < 0.01.