Figure 4: Delayed recovery of mitochondrial filamentous morphology in SAD fibroblasts. (a) Representative confocal microscopy images showing the mitochondrial morphology of healthy and SAD fibroblasts untreated (control), after 7 h with 20 μM CCCP (total CCCP) or after 6 h of CCCP treatment followed by 1 h without it (reversion). (b)–(d) Quantification of mitochondrial morphology of healthy-SAD couples when the cells remain untreated (b), treated with CCCP for 7 h (c), or reversibly treated with CCCP (d). (e) Mitochondrial membrane potential measured by TMRM of fibroblasts treated as in (a). Mitochondrial morphology classification according the length of the branches in filamentous (branch > 2.3 μm), fragmented (branch < 1.8 μm), and intermediate (1.8 ≥ branch ≥ 2.3 μm) patterns. different healthy/AD sex- and age-matched couple samples: AG11362/AG05809, AG07310/AG06869, and AG11362/AG05809. Scale bar: 40 μm. and ; ns: not significant.