Fig. 2
From: Connecting lysosomes and mitochondria – a novel role for lipid metabolism in cancer cell death
Lysosomal stress alters lipid metabolism. HUH7 cells were treated as indicated (24 h). Relative mRNA expression levels of PGC1α (a) and PPARα (b) were detected by qPCR. c Protein expression of PGC1α and PPARα was detected by WB. Total protein served as loading control. d CD36 expression was visualized by antibody staining and determined by flow cytometry. e Levels of free fatty acids were determined by a coupled enzyme assay, which results in a colorimetric product proportional to the fatty acids present (MAK044 Sigma Aldrich). Absorbance was quantified relative to DMSO control. f, g Cardiolipin content was analyzed using a fluorimetric detection kit (K944 Biovision) in whole cell lysate (f) and in isolated mitochondria (g). h Cells were labeled with Bodipy 558/568 Red C-12 (cyan). Cox4 and Lamp3 (magenta) were visualized by antibody staining and nuclei (yellow) by Hoechst 33342, respectively. Cells were analyzed by confocal microscopy. Scale bar 7.5 μm. Representative images out of three independent experiments are shown. Bars are the mean + SEM of three independent experiments. p* < 0.05 (One-way ANOVA, Dunnett post test)