Fig. 1: Siah2 redistribution on FCCP-depolarized mitochondria.
A ΔΨm in cortical neurons treated with 10 μM FCCP for 1 h and a its relative quantification. ΔΨm was measured by using the fluorescent probe TMRE and confocal microscopy. The reduction of the intensity of TMRE fluorescence (red) is indicative of mitochondria depolarization. The green fluorescent signal corresponds to cells loaded with Fluo4-AM used to identify neuronal morphology in control and FCCP treatment conditions. a Quantification of the results reported in A; each bar represents the mean±S.E.M of the percentage of different experimental values obtained in three independent experimental sessions. *P < 0.05 vs CTL. B Siah2 expression was measured by western blotting in, cytosolic, and mitochondrial fractions of cortical neurons exposed to 1 h FCCP. Immunoblotting for VDAC and Tubulin was used to define the identity of mitochondrial and cytosolic compartments, respectively. C Left panel: Siah2 protein expression in cortical neurons after FCCP treatment. Each bar represents the mean±S.E.M of the percentage of different experimental values obtained in three independent experimental sessions. *P < 0.05 vs CTL. Right panel: representative cortical neuron double-labeled with Siah2 (green) and Mito-RFP (red). Siah2/Mito-RFP colocalization was 100% in CTL conditions and was up to 133% ± 1 after FCCP treatment. D Left panel: LC3-II protein expression in cortical neurons after FCCP treatment. Each bar represents the mean±S.E.M of the percentage of different experimental values obtained in three independent experimental sessions. Right panel: representative cortical neuron double-labeled with LC3-GFP (green) and Mito-RFP (red). LC3-GFP/Mito-RFP colocalization was 100% in CTL conditions and was up to 154% ± 1 after FCCP treatment. Bars 10 μM.
