Figure 3
From: Sirtuin 5 depletion impairs mitochondrial function in human proximal tubular epithelial cells
SIRT5-knockdown disrupts the mitochondrial fission/fusion machinery. hPTECs were transfected using siRNA against SIRT5 or non-targeting siRNA control. Seventy-two hours post-transfection cells were exposed to 6 h of OND (1%O2 + HBSS) or control conditions [normoxia and complete medium (21%O2 + CM)]. (A) Representative WBs of pro-fission proteins DRP1, DRP1-S616, the DRP1 receptors mitochondrial MiD51 and FIS1. Scatter plots showing densitometry results of DRP1, DRP1-S616, MiD51 and FIS1. DRP1 and DRP1-S616 were normalized to tubulin while MiD51 and FIS1 where normalized to actin. Whole WBs shown in supplementary file (Supplementary Fig. S8c,d). (B) WBs of pro-fusion proteins MFN1/2 and OPA1. Scatter plots showing densitometric quantitation of MFN1, MFN2 and OPA1. Data are from three to four independent experiments with n = 3 replicates/group. MFN1 and OPA1 were normalized to tubulin while MFN2 was normalized to actin. Whole WBs shown in supplementary file (Supplementary Fig. S8e). ImageJ was used for densitometric analysis. To determine statistical significance a two-way ANOVA was carried out followed by Tukey’s post hoc test to normalize for multiple comparisons. Data are mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001.
