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. 2022 Nov 25;14(23):5812. doi: 10.3390/cancers14235812

Figure 3.

Figure 3

Elaiophylin inhibits mitophagy and induces mitochondrial dysfunction in A549 cells. A549 cells were exposed to elaiophylin or FCCP (a mitophagy agonist) for 24 h. (A) The expression of autophagy-related proteins (LC3B and SQSTM1 (p62)) was assessed by Western blot analysis. (B) The expressions of mitophagy-related proteins (PINK1 and Parkin) were assessed by Western blot analysis. (C,D) A549 cells overexpressing Mito-Keima plasmid were treated with elaiophylin for 24 h. Mito-Keima (red fluorescence) was detected by a fluorescence microscope. FCCP, a mitophagy agonist, was used as a positive control. (E) Co-localization of mitochondria and lysosomes was assessed by MitoTracker (200 nM) and LAMP1 co-staining. (F) Mitochondrial membrane potential was assessed by rhodamine 123 staining. Data were expressed as means ± SD of three experiments and each experiment included triplicated repeats. * p < 0.05, ** p < 0.01 vs. control. Original Western Blots can be found at supplementary materials.