Fig. 3

QW24 degraded BMI-1 protein through autophagy-lysosome pathway. a, HCT116 cells were pretreated with proteasome inhibitor MG132 (10 μM) or lysosomal inhibitor Chloroquine (100 μM) for 2 h as indicated, and then 2 μM QW24 was added to cells for 6 h. Cells were lysed and BMI-1 protein level was measured by western blotting analysis. b, HCT116 cells were treated with indicated concentrations of QW24 for 12 h and LC3 (the autophagy marker) protein level was measured by western blotting analysis. c, d, HCT116 cells were treated with indicated concentrations of QW24 for 12 h (c) or 2 μM QW24 for indicated time (d). Then cells were lysed and LC3 protein level was measured by western blotting analysis. e and f, HCT116 cells were transfected with GFP-LC3 plasmid and treated with 2 μM QW24. Cells were imaged by fluorescence microscope at the indicated time (e), and the punctate cytoplasmic LC3 distribution cells were counted (f). Scale bars, 10 μm. Data are presented as mean ± s.d. (n = 5); n.s., Not statistically significant; ***, P < 0.001