Fig. 8 |. Adipose Ndufv2 overexpression increased mitochondrial biogenesis and protein levels via RoS generation by altering supercomplex composition in a strain-by-sex manner.

Comparisons of a, MTDR staining of the mitochondria and b, frozen respirometry quantifications in frozen gonadal adipose homogenates between GFP and NDUFV2 overexpressing female A/J mice (GFP n = 6; NDUFV2 n = 8). Similarly, c, MTDR staining of the mitochondria and d, frozen respirometry quantifications were measured between GFP and NDUFV2 overexpressing male A/J mice (GFP n = 7; NDUFV2 n = 8). Likewise, e, frozen respirometry quantifications and f, immunoblot analyses of CPT1a, LCAD, UCP1 and vinculin (loading control) levels and their corresponding quantifications from female C57BL/6 J mice (n = 7 per group except UCP1 n = 4 per group) and g, frozen respirometry quantifications and h, immunoblot analyses of CPT1a, LCAD, UCP1 and vinculin (loading control) levels and their corresponding quantifications from male C57BL/6 J mice (n = 8 per group except UCP1 n = 4 per group), respectively, are shown. Finally, i, immunoblot staining of NDUFV2 overexpression; j, oxygen consumption rates before and after the addition of N-acetylcysteine (NAC) (n = 10 replicates per group), and k, blue native gel electrophoreses followed by immunoblotting for NDUFA9 (complex I), Core2 (complex III) and SDHA (complex II) for supercomplex visualization in stably transduced differentiated 3T3-L1 cells are shown. Data are presented as mean ± s.e.m. P values were calculated using two-factor ANOVA corrected by post-hoc Holm–Sidak’s multiple comparisons test (b, d, h, j) or unpaired two-tailed Student’s t-test (a, c, j). See also Extended Data Fig. 10 and Supplementary Fig. 3.