Figure 5.

SIRT5-depletion exacerbates OND-induced bioenergetic dysfunction. hPTECs were transfected with siRNA against SIRT5 or non-targeting siRNA control. Seventy-two hours post-transfection cells were exposed to 6 h of oxygen and nutrient deprivation (OND) (1%O₂ + HBSS) or normoxia and complete medium (21%O₂ + CM). (A) TEM images showing mitochondria (red arrows). Magnification: ×1500 and ×8000. Lower panels show higher magnification of the area indicated by blue squares. (B) Summary of Seahorse XFp Mito Stress Test assay of cells exposed to 6 h OND. Representative profile of Mito Stress Test data showing oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) with arrows indicating injection of the specific stressors oligomycin (O), FCCP (F) and Rotenone plus antimycin A (R + A). Relative values of the parameters normalized to the non-targeting siRNA control. Data are from four independent experiments with n = 3 technical replicates/group. Wave software (Agilent) was used for data analyses. Mann–Whitney U test was used to determine statistical significance. Data are median ± IQR. *p < 0.05.