Figure 5.

Association of vMIA with adenine nucleotide translocator (ANT). (a) Lysates from cells constitutively expressing myc-tagged vMIA, or from control cells (pcDNA3 transfected) were immunoprecipitated with the anti-myc antibody 9E10 covalently linked to Affi-Prep-10 beads. Bound proteins were separated by SDS/PAGE and stained with colloidal Coomassie blue (Novex). The bands designated vMIA and ANT were identified by microsequencing. Asterisks indicate nonspecific bands. (b) (Upper) 293T cells were transiently cotransfected as indicated with expression plasmids (0.5 μg each) coding for vMIAmyc, V5-tagged ANT-1, V5-tagged β-gal, or the empty vector. vMIA was then immunoprecipitated (lanes designated IP) with anti-myc antibody and interacting proteins were detected by Western blot analysis with anti-V5 antibody (Invitrogen). Lanes designated L are samples of the transfected cell lysate taken prior to immunoprecipitation. Asterisks indicate bands of antibody heavy and light chains. (Lower) The blot was reprobed with anti-myc to confirm expression of vMIA-myc.