Fig. 1: Opa1 determines nucleoid abundance and array in the mitochondrial population and intramitochondrial longitudinal distribution.
From: OPA1 and disease-causing mutants perturb mitochondrial nucleoid distribution
A Mitochondrial DNA abundance in WT MEF, Opa1−/− and Opa1−/− cells stably expressing a lentiviral plasmid carrying OPA1 cDNA, quantified by qPCR and expressed as mt-Nd4 to Gapdh ratio. Data are mean ± SEM of ≥ 3 independent experiments. B Schematic representation of the components of nucleoid distribution. C Representative images of WT MEF, Opa1−/− and Opa1−/− cells with WT OPA1 acute expression, co-transfected with mCherry-Omp25 (OMM) and Tfam-GFP (nucleoids) cDNA. Bottom panel, insets of Tfam-GFP (top) and merge (bottom). D Percentage of mitochondria with different number of nucleoid clusters. E Mean nucleoid cluster number per mitochondrion. F Mean nucleoid cluster number per mitochondrion area, manually quantified. G Number of nucleoid clusters per total mitochondrial area, semi-automatically quantified by the MiNuD algorithm. For Opa1−/− cells, DNA was labeled using PicoGreen, and mitochondria were labeled with mCherry-Omp25 (OMM) or MitoTracker Deep Red. H Mean nucleoid cluster number per mitochondrion length. I Simple regression of the relationship between nucleoid cluster number and mitochondrion length in individual organelles. Data are mean ± SEM of ≥ 281 objects from ≥ 15 cells of ≥ 3 independent experiments, and for G data are mean ± SEM of ≥ 10 cells of ≥ 2 independent experiments (****p < 0.0001; ***p < 0.0005; *p < 0.05).
