Fig. 2: Parkin translocation to mitochondria is regulated by Calcineurin.
A Representative confocal images of MEF cells transfected with mCherry-Parkin, mito-YFP and dominant negative CaN (ΔCnAH151Q) or the empty vector (EV) for 2 days before being treated with DMSO or 10 μM CCCP for 3 h. B Quantification of A. Graph bar shows mean ± SEM of percentage of cells with mCherry-Parkin on mitochondria for at least ≥ 300 cells per biological replicate. Two-way ANOVA followed by Tukey’s multiple comparison test (n = 3-9; p < 0.0001). C Quantification of A using Squassh. The graph bars show mean ± SEM of Squassh colocalization coefficient for at least ≥ 50 images per biological replicate. 0 = no colocalization, 1 = perfect colocalization. Two-way ANOVA followed by Tukey’s multiple comparison test (n = 4-5; p < 0.001). D Representative confocal images of MEF cells transfected with mCherry-Parkin, mito-YFP in which CaN was downregulated, and relative control. E Quantification of D. Graph bar shows mean ± SEM of percentage of cells with mCherry-Parkin on mitochondria for at least ≥ 300 cells per biological replicate. Two-way ANOVA followed by Tukey’s multiple comparison test (n = 3; p < 0.01). F Quantification of D using Squassh. The graph bar shows mean ± SEM of Squassh colocalization coefficient for at least ≥ 50 images per biological replicate. 0=no colocalization, 1 = perfect colocalization. At least 3 independent experiments were performed. Two-way ANOVA followed by Tukey’s multiple comparison test (n = 3; p < 0.01).
