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Fig. 2. Antibody characterization by subcellular fractionation and western-blot analysis. HeLa and 143B cells (normal {rho}+ and mtDNA-less {rho}0) were subjected to subcellular fractionation. Equal protein amounts of total homogenate (TH), mitochondrial pellet (MP) and post-mitochondrial supernatant (PMS) were separated by SDS-PAGE and transferred to membranes. Arrowheads point to the positions of marker proteins with the indicated molecular mass (kDa). The distributions of outer membrane Mfn2 and inner membrane COX2 demonstrate mitochondrial enrichment in both cell types. The {rho}0 cells do not have any mtDNA-encoded COX2. The mtTFA-protein has an apparent molecular mass of 25 kDa and is strongly downregulated in {rho}0 cells. The mtSSB-protein has an apparent molecular mass of 16 kDa and is present at similar levels in fractions of {rho}+ and {rho}0 cells.





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