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. 2016 Feb 11;11(2):e0148213. doi: 10.1371/journal.pone.0148213

Fig 2. ADCK3 binds members of the CoQ biosynthetic complex in vitro.

Fig 2

(A). ADCK3 interacts with Coq3, 5, 7 and 9 in vitro. Recombinant GST-Coq proteins were expressed and purified (left panel–Collodial Coomassie Staining) prior to use in pull down assays with mitochondrial extracts from HeLa cells (right panel). Ponceau staining was performed to identify GST-Coq proteins and their concentration. Immunoblotting was also performed with anti-ADCK3 and anti-Tom20 antibodies. (B, C). BN-PAGE data from Triton-X100 (B) and Digitonin (C) solubilised mitochondria from HEK293 cells. 50 μg mitochondria were solubilised and separated via 2D-BN-PAGE. Immunoblotting performed with antibodies to ADCK3 and Core I (Complex III component). 1D SDS-PAGE of mitochondrial extracts was performed as a control for antibody staining (see panels to left of 2D gels).