Determination of Dox-induced OXPHOS protein levels in healthy and Barth lymphocytes.

<p>(A) Representative western blot for the detection of OXPHOS proteins in healthy and Barth lymphocytes. Healthy and Barth lymphocytes were treated with 0.1, 0.5, 1, and 5 μM of Dox for 24 h and OXPHOS protein levels were determined by western blot using an antibody cocktail containing five mouse mAbs against mitochondrial complex subunits I (NDUF8-20 kDa)), II (SDHB-30 kDa), III (UQCRC2-48 kDa), IV (MTCO1-40 kDa) and V (ATP5a-55 kDa). Actin was used as a loading control to determine the relative levels of each subunit. (B-F) Quantitative determination of mitochondrial complex subunits from (A) normalized by β-actin. In each panel, basal level of mitochondrial complex subunits for control (untreated) healthy and Barth lymphocytes were normalized to 100; and Dox-induced changes in the band intensities of each subunit in the western blot were normalized against corresponding control.