Fig. 2

Blue Native PAGE and Western blot analysis of patient fibroblasts and liver. a The presence of respiratory chain complexes I-IV (CI-CIV), CIII assembly and BCS1L protein from the patient (P) and controls (C1-C3) were analyzed in fibroblast mitochondria using BN PAGE technique. C1 and C2 are fibroblasts from umbilical cords from healthy pregnancies, C3 are fibroblasts from a child with no symptoms of mitochondrial disease. Monomers (lower band) and oligomers (upper band) of BCS1L were detected using antibodies raised against this protein. CIII was investigated using antibodies directed against the two CIII subunits RISP (mature CIII) and CORE1 (lower band pre-CIII, upper band mature CIII). CI was assessed using an antibody against NDUFV1. Antibodies against 30 kDa IP and cytochrome c oxidase subunit Va (COXVa) were used to detect CII and CIV, respectively. The data shows a clear reduction of mature CIII complexes (with incorporated RISP) in the patient cells and loss of BCS1L protein. The amount of the other complexes (CI, CII and CIV) in patient cells and C3 is less than in C1 and C2, but the ratios of the individual complexes are similar in-between the samples. b Western blot analysis of homogenates from liver and fibroblasts from the patient (P) and two controls (C1 and C2). A loss of BCS1L protein and clear reduction in liver RISP is seen in accordance with BCS1L deficiency