Figure 6

CBD interacts with purified VDAC1 to decrease its channel conductance. (a) VDAC1 was reconstituted into PLB and channel currents in response to voltage steps, 0 to −10 mV (I) and 0 to −60 mV (II), were recorded before (control) and 15 min after the addition of 20 μM CBD. A representative of three independent experiments is shown. The dashed lines indicate zero current. (b) The effect of CBD on VDAC1 conductance as a function of voltage, from 60 mV to −60 mV. The average steady-state conductance at any given voltage was normalized to the conductance at 10 mV. The recordings were taken before (control) and 15 min after the addition of CBD. (c) Thermophoretic analysis of CBD–VDAC1 interaction. The interaction between the fluorescently labeled purified VDAC1 and CBD (or the endocannabinoid AEA) was tested using thermophoresis (Laser power 80%). VDAC1 and CBD (or AEA) were incubated for 30–60 min in PBS, with 2% ethanol as vehicle. The difference in fluorescence is plotted against CBD or AEA concentration. The data represent an average of three independent experiments (two of them in duplicates) for CBD, and two independent experiments in duplicates for AEA