Fig. 6

Cell-type-specific effect of CAMKK2 loss-of-function on the SDH and OXPHOS-associated MPCs. A, B, E, F Immunoblots showing OXPHOS-associated MPCs in the parental and CAMKK2^−/− HEK293 cell clone-derived mitochondria. The immunoblot was generated by simultaneous use of a cocktail of six antibodies (Table 1). C, G Immunoblots in AB and EF were false colored and overlaid to show relative amount\shift of OXPHOS-associated protein complexes. Vertical alignment of OXPHOS-associated proteins indicates association in multiprotein complexes that co-migrated during first-dimension native PAGE. D Immunoblots showing relative abundance of SDHA, SDHB and SDHC in complex II. The vertical alignment of SDHA, SDHB and SDHC indicates their association in a single megacomplex. Interpretation of colored markings and arrows: A–C Red/pink rectangles: potential megacomplex involving different OXPHOS complexes. White dotted rectangle: a > 1200 kDa MPCs potentially associated with different OXPHOS complexes to form respirasome structures. E–G Blue rectangles: differential oligomerization of ATP5A and UQCRC2. Green rectangles: differential shift of MT-CO2 MPCs. Red rectangle: differential shift of ATP5A. White rectangle: vertical alignment of SDHB MPCs. White and yellow arrow: relative abundance or differential shift of respective MPCs. D Grey arrows: placement of excised first dimensional BN-PAGE gel on the second dimension SDS-PAGE. Yellow arrow: Incubation of the same blot with a different antibody without stripping. Blue dotted rectangle: relative abundance of SDHB