fig7

Figure 7. Treatment with rapamycin or MitoTEMPO decreases oxidative stress and enhances mitophagy and OCR in old age LCR rat VSMCs. Representative confocal images of VSMCs treated with and without 20 nM rapamycin (A), or MitoTEMPO (B) and co-stained for lysosomal marker LAMP1 (green) and mitochondrial marker TOM20 (red). Bright yellow fluorescence indicates colocalization of LAMP1 and TOM20 and active mitophagy. Data are presented as average integrated density/ROI ± SEM where n = 4. Scale is 10 μm. Representative confocal images of mitochondrial ROS levels with and without rapamycin (C) or MitoTEMPO (D) treatments as measured by MitoSOX Red (MSR) fluorescence. Intensity of fluorescence was presented as integrated density (mean ± SEM, n = 4, Scale is 10 μm). OCR was measured at the basal level and in the presence of 2 μM oligomycin, 500 nM trifluoromethoxy carbonylcyanide phenylhydrazone (FCCP), and 0.5 μM antimycin A + 500 nM rotenone in VMSCs of old age LCR rats treated without and with rapamycin (E and G) or MitoTEMPO (F and H). Mitochondrial bioenergetic parameters were derived from VSMC OCR measurements.