Fig. 2: SIM2 expression levels alter MRC complex and supercomplex (SC) formation and activity.
From: Noncanonical role of singleminded-2s in mitochondrial respiratory chain formation in breast cancer
Mitochondrial proteins shown in a, e, i are from SIM2KO MCF7 cells, and and those shown in b, f, j are from SIM2s-FLAG SUM159 cells. All cells were solubilized with 1:8 protein:digitonin, and the proteins were subjected to BN-PAGE followed by immunoblot analysis to measure the levels of a, b NDUFB8, e, f MT-CO2, and i, j UQCRC2. Mitochondrial proteins from c, g SIM2KO MCF7 cells and d, h SIM2s-FLAG SUM159 cells were solubilized with 1:8 protein:digitonin and subjected to CN-PAGE followed by an in-gel activity assay of c, d Complex I visualized by NTB oxidation and g, h Complex IV visualized by DAB oxidation. k, l SDS‒PAGE was followed by immunoblotting for measuring the level of the mitochondrial protein TOM70 in k SIM2KO MCF7 cells and l SIM2s-FLAG SUM159 cells. m Malformation of CIII inhibits electron flow through the MRC.
