Figure 3. New synthesized compounds bind to TSPO and induce Δψm collapse.

(A) [³H]PK11195 radioligand binding: membranes homogenates obtained from kidney (20 μg of proteins) were incubated with 0.6 nM [³H]PK11195 and different compounds concentration. Reaching equilibrium, samples were filtered and bound radioactivity was counted. Data are expressed as percentage of specific binding versus basal value (set to 100%) and represent the mean ± SEM of three different experiments. (B) Evaluation of Δψm in isolated mitochondria: Mitochondria (5 μg of proteins) were treated with compound 1 (5 μM), or compound 7 (5 μM), or DMSO (control) for 15 min. The Δψm was evaluated using JC-1 protocol as describe in Methods. The data are expressed as variation of JC1 uptake into mitochondria, calculated as the difference between RFU read after 10 minutes and RFU read at the beginning. Data represent the mean ± SEM of three different experiments. Each experiment was performed in duplicate. Statistical significance was determined with a one-way ANOVA with Bonferroni post-test:**P < 0.01, ***P < 0.001 vs control. (C) Representative graph of mitochondria potential evaluation using JC-1 protocol. The results were expressed as RFU units in time.