Autolysosomes were not formed by conessine treatment.

(A) Autophagosome formation by conessine. HEK293 cells were transfected with a plasmid encoding mRFP-GFP-LC3. After 24h, the transfected cells were incubated with the indicated concentrations of conessine (0, 2.5, 5, 10, and 20 μM) for 24 h. Cells were analyzed with confocal microscopy. (B) p62 accumulated in autophagosomes after conessine treatment. HEK293 cells were transfected with a plasmid encoding GFP-LC3, and the cells were treated with indicated concentrations of conessine, followed by immunostaining with anti-p62 antibody. Cells were analyzed with confocal microscopy.