Fig. 6: Leptin initiated OMA1 disruption through ubiquitin/proteasome-mediated mechanism.
a, b OMA1 and YME1L were detected by western blot for both hMSCs-Leppre and hMSCs-Ctrlpre cultured in GSDH for 24 h, and their protein levels were quantified by normalization to β-actin. c, d OMA1 protein level was quantified by western blot after hMSCs were treated with CHX (10 μg/ml) for the indicated duration using β-actin as a loading control, the OMA1 expression levels just before of CHX treatment were set as a baseline level. e, f Western blot of OMA1 and OPA1 isoforms protein levels of hMSCs-Leppre and hMSCs-Ctrlpre were co-incubated with MG132 (2 μM), SB216763 (10 μM), or CQ (50 nM), respectively, in GSDH culture for 24 h and quantified in bar graphs. g HEK293T cells were transfected with Flag-OMA1 and HA-ubiquitin (HA-Ub) or empty vector for 48 h, and then, treated with MG132 (10 μM) for an additional 2 h; leptin (50 ng/ml) was administered throughout the whole period. WCLs harvested from these cells were explored by IP with anti-HA affinity gel; subsequently, Flag-OMA1 protein level was measured by western blot using anti-Flag antibody
