Figure 3
From: Piperlongumine induces autophagy by targeting p38 signaling
PL enhances autophagy formation, does not block autophagy flux. U2OS/GFP-LC3 cells were pre-treated with 1 mM 3-MA, 10 μM CQ, or 10 nM Baf-A1 for 6 h. zVAD (50 μM) and PL (5 μM) were then added. The cells were collected for the analysis post 16-h treatment. (a) Punctate GFP-LC3 fluorescence in U2OS/GFP-LC3 cells treated with PL in the presence or absence of various autophagy inhibitors. (b) The average numbers of punctate GFP-LC3 fluorescence dots were calculated in 200 cells. *P<0.001. (c) Expression of ATG5, Beclin-1, and GFP-LC3 in cells treated with autophagy inhibitors in (a and b) were determined with western blot. Thirty micrograms of WCE proteins were loaded. GAPDH served as a loading control
