Figure 6
From: Pomegranate activates TFEB to promote autophagy-lysosomal fitness and mitophagy
PE reduces damaged mitochondria and alleviates mitochondrial redox toxicity. (a) Top: Fluorescence images of SY5Y cells stably expressing MitoTimer reporter treated with DMSO or 10 μM CCCP in the absence or presence of 300 μg/ml PE for 16 h. Smaller inlets show the green (young mitochondria) and red (aged and damaged mitochondria) MitoTimer expression under each condition. Bottom: Quantification of the average red:green intensity ratio of MitoTimer per cell. (b) Top: Measurement of mitochondrial ROS via Mitosox staining and TOMM20 immunofluorescence in SY5Y cells treated with DMSO, 10 μM CCCP alone or 10 μM CCCP in the presence of 300 μg/ml PE or 300 μg/ml PE supplemented with lysosomal inhibitor (NL) for 16 h. Bottom: Quantification of Mitosox intensity level per cell area, expressed as fold change against DMSO. At least 30 cells from random fields were analyzed for MitoTimer and Mitosox assays. Nuclei were stained with DAPI. All values are mean + S.E.M (n = 3–7). Differences against DMSO, CCCP or CCCP + PE are significant at *p < 0.05, ***p < 0.005 and ###p < 0.005. Scale bar, 10 μm.
