Figure 4
From: Activation of lysosomal mediated cell death in the course of autophagy by mTORC1 inhibitor
SDS-203 initiates incomplete autophagy by inhibiting autophagy flux. (A, B) MIA PaCa-2 cells were treated with SDS-203 (20 μM) for 6, 12 and 24 h in presence or absence of bafilomycin A1 (80 nM) or NH4Cl (10 mM) and LC3 turnover was detected and quantified by western blot analysis. (C, D) SDS-203 (20 µM) treated MIA PaCa-2 cells were allowed to grow in the presence or absence of rapamycin (200 nM, 12 h) or starved for (12 h), followed by protein expression examination of LC3-II and p62. (E–G) MIA PaCa-2 cells were incubated with SDS-203 in the presence or absence of rapamycin or starvation and the effects on LC3-II expression was visualized using confocal microscopy, LC3-II puncta quantification was done by using ImageJ software; the panel represents the set of images of at least three independent biological replicates. Statistics, Student’s t test *p < 0.05, **or##p < 0.01, ***p < 0.001 vs. control (0.1% DMSO).
