Figure 6.

Activation of SIRT1/Nrf2 signaling attenuates the anti-mitophagy effect of elaiophylin in A549 cells. The cells were divided into six groups: control, SIRT1 overexpression only, Nrf2 overexpression only, elaiophylin, elaiophylin+SIRT1 activation (SIRT1 overexpression plus SRT1720), elaiophylin+Nrf2 activation (Nrf2 overexpression plus DMF). (A) The expressions of mitophagy-related proteins PINK1 and Parkin were assessed by Western blot analysis. (B) Cells overexpressing Mito-Keima were treated with elaiophylin for 24 h. Mito-Keima (red fluorescence) was assessed by a fluorescence microscope. FCCP, a mitophagy stimulant, was used as a positive control. (C) Colocalization of mitochondria and lysosomes was assessed by MitoTracker (200 nM) and LAMP1 staining. Data were expressed as mean ± SD of three experiments. ** p < 0.01 vs. control, ^## p < 0.01 vs. elaiophylin group. Original Western Blots can be found at supplementary materials.