Figure 7
From: Activation of lysosomal mediated cell death in the course of autophagy by mTORC1 inhibitor
TFEB translocation and lysosomal enzyme activation upon suppression of mTORC1 by SDS-203 treatment. (A, B) Western blot representation and band quantification of TFEB protein expression in cytosolic and nuclear fraction of MIA PaCA-2 samples treated with SDS-203 (0, 10, 20 µM) for 24 h. (C) confocal image representation showing TFEB nuclear translocation upon SDS-203 treatment (0, 10, 20 µM) for 24 h in MIA PaCa-2 cells(D, E) Western blot illustration of TFEB knockdown i n MIA PaCa-2 cells and its densitometric quantification. (F, G) shows western blot for LAMP1 expression upon SDS-203 treatment in TFEB Knockdown (siRNA) and TFEB scramble g MIA PaCa-2 cells and its densitometric quantification (H) Effect of SDS-203 on lysosomal activity: MIA PaCa-2 cells were treated with SDS-203 (20 µM) with or without NH4Cl, cell lysates were then subjected to measure lysosomal enzyme activity by analyzing rhodamin110 fluorescence. (I) Shows time course western blotting analysis and (J) densitometric quantification of lysosomal enzymes- cathepsin D/L in MIA PaCa-2 cells upon SDS-203 treatments. Protein fold change was quantified by using ImageJ software. Error bar indicates ± SEM. *p < 0.05, **p < 0.01, ***or###p < 0.001 vs. control.
