Research Paper Volume 12, Issue 8 pp 6700—6732
Mitochondrial nucleoid remodeling and biogenesis are regulated by the p53-p21WAF1-PKCζ pathway in p16INK4a-silenced cells
Figure 3. Mitochondria are activated in senescent fs-HDF cells. To confirm the role of PKCζ in the regulation of mitochondrial nucleoid remodeling, fs-HDF young cells were treated with doxorubicin (Doxo; 100 ng/mL) for 7 days and senescence was monitored. (J) Doxo-treated young cells were transfected with siControl or siPKCζ for 48 h and subjected to RT-qPCR analysis. Note the effect of PKCζ knockdown on mitochondrial biogenesis. (K) Fluorescence microscopy of mitochondrial nucleoid remodeling in Doxo-induced senescent cells. BrdU incorporation (red) in enlarged mitochondria was lost in the siPKCζ-transfected cells. Scale bars, 20 μm. (L) BrdU incorporation in mitochondria was quantified. Fluorescence microscope images were captured and counted at least 200 cells using ImageJ software (n=10 images/group). Data are means ± SD of three independent experiments per group. One-way ANOVA followed by Tukey HSD post hoc test.