FIGURE 1.

Immunochemical analyses of rat liver mitochondrial fractions with antibodies against mitochondrial and nonmitochondrial markers. Proteins collected during mitochondrial isolation and purification steps (M2, 10,000 × g; M4, 10,000 × g; M5, 9,000 × g; MT, intact MT after repeated differential centrifugation and Percoll gradient purification; and CO, control tissue homogenate) were separated by SDS-PAGE and transferred onto nitrocellulose membranes as described (see under “Experimental Procedures”). Western blotting was performed with antibodies against the following: A, mitochondrial matrix marker, GRP 75; B, mitochondrial outer membrane marker, VDAC; or C, cytosol marker, tubulin. The immunocomplexes were visualized by the HRP-conjugate reaction using a chemiluminescence substrate (Pierce). MT lane, run on the same gel as other fractions, was cropped and pasted alongside other fractions after removing the intervening lanes.