Fig 4.
Assembly of Sym1 into mature complexes. (A) Yeast mitochondria were solubilized in digitonin buffer, separated by BN-PAGE, and analyzed by Western blotting using the PAP antibody; (B) Sym1ZZ isolated from mitochondria by IgG chromatography was analyzed by BN-PAGE and stained with Coomassie brilliant blue; (C) mitochondria were solubilized in digitonin-containing buffer, and complexes were separated by 1st-dimension BN-PAGE and 2nd-dimension SDS-PAGE and analyzed by Western blotting with the anti-Sym1 antibody; (D) mitochondria were solubilized in buffers containing digitonin (Dig.) or dodecyl maltoside (DDM), and the complexes were separated by 1st-dimension BN-PAGE and 2nd-dimension SDS gel and analyzed by Western blotting; (E) Sym1 import was monitored with BN-PAGE and subsequent digital autoradiography; (F) import and BN-PAGE analysis of truncation constructs described for Fig. 3D. Complexes were visualized by digital autoradiography.