VDAC, ANT and TSPO detection in human red blood cell ghosts. Samples (15 μg of protein) of whole lysates were subjected to SDS-PAGE (10% acrylamide) and stained with Coomassie blue (A) or analyzed by Western blotting using polyclonal anti-ANT (1:1000 dilution), rabbit polyclonal anti VDAC1 -2, -3 (1:100 dilution) or polyclonal goat anti-TSPO raised against the C terminus of human TSPO (1:1000). The positions of molecular weight (kDa) protein standards are indicated by arrows. (B) 4 bands appearing between 29-30 kDa and 36 kDa correspond to different isoforms of VDAC and marked band at 58 kDa correspond to dimers of VDAC1 isoform (29-30 kDa). Multiple bands at higher molecular weights suggest oligomerization of VDAC proteins. ANT and TSPO proteins are also clearly visible. It is to be noted that, according to the supplier assessment (Everest Biotech), the TSPO protein could not be expected at 18 kDa but rather at 36 kDa. These blots are representative of 12 replicates. Immunofluorescence experiments (C) were performed on smears prepared as described in “Immunostaining.” Dilution were 1/5 for primary and 1/20 for secondary antibodies. Scale bars represent 10μm.