Figure 5.

VDAC polymerization is induced by TSPO ligands. Red blood cells (RBCs) were incubated 60 min at 37 °C in 3 different culture media: control (MKB), low cAMP (1 µM 2-Me-S-ADP) and Ca²⁺-depleted (without Ca²⁺). Then they were incubated 10 min at 37 °C in the presence of TSPO ligands Ro5-4864 or NCS1018. Ghost membrane were prepared from each treatment, lysed with 1% Triton X-100 and analysed using an anti-human VDAC1 polyclonal antibody. Monomer, dimer, trimer and tetramer bands of VDAC were identified (Panel A). Anti-b actin antibody was use as loading control (Panel B). Anti-ANT antibody immunolabeling showed only a trimeric band (Panel C). Densities of VDAC immunogenic bands were quantified to calculate dimer/monomer (Panel D) and trimer/monomer (panel E) ratios, under control (MKB, white bars) or experimental conditions (2-Me-S-ADP, grey bars and medium without Ca²⁺, black bars). Results are expressed as means ± SEM of the polymer/monomer ratios, and considered different from basal (control, unstimulated) values when *p < 0.05; n = 4.