Figure 1
From: Sirtuin 5 depletion impairs mitochondrial function in human proximal tubular epithelial cells
SIRT5 expression increases after ischemia in PTECs in vivo and in vitro. (A,B) Male C57BL/6J mice underwent either right uninephrectomy followed by clamping of the left renal pedicle for 30 min (IRI group; bilateral IRI) or sham surgery. Tissues were harvested 48 h post-surgery. (A) IHC analysis. Formalin-fixed, paraffin-embedded (FFPE) kidneys (sham and IRI) were screened for SIRT5. Rabbit IgG at an equivalent concentration served as a control. (B) Immunofluorescence analysis. Murine FFPE kidney sections (sham and IRI) were co-stained for SIRT5 (red) and LTL (green), a marker of PTECs. 4′,6-diamidino-2-phenylindole (DAPI) (blue) was used to visualize nuclei. Dashed lines in the sham-surgery group define low SIRT5 expression in PTs. G: Glomeruli. Scale bars: 100 µm. n = 3–5 mice/group. (C,D) hPTECs were exposed to control conditions (21%O2 + CM) or OND (1%O2 + HBSS). (C) Bar graph showing SIRT5 mRNA expression. (D) Representative WBs and scatter plot of densitometric analysis showing SIRT5 protein levels. Whole WB shown in supplementary file (Supplementary Fig. S7a). Data are from four independent experiments with n = 3 replicates/group. mRNA levels were normalized to HPRT1. SIRT5 protein levels were normalized to actin. ImageJ was used for densitometry. To determine statistical significance of mRNA and protein data, a Mann–Whitney test was carried out. Data are median ± IQR. *p < 0.05.
