Fig 1: Defective mitochondrial import upon expression of mutant renin isoforms. (A) Representative western blot analysis showing Mito-YFP when co-expressed with the indicated renin isoforms. ‘Val’ indicates cells treated with valinomycin as a positive control of defective mitochondrial import. (B) Quantification of Mito-YFP import, measured as the ratio between precursor (YFP with the MTS, upper band) and mature (YFP without the MTS, lower band) forms. **P<0.01; ***P<0.001; ****P<0.0001; two-way ANOVA with Bonferroni post hoc test versus WT. Data are shown as mean±s.d. (n=4-14 independent experiments per group). (C) Immunofluorescence analysis showing merged images of the indicated renin construct (green), a mitochondrial marker (TIM44, in red) and nuclei (DAPI, blue). Yellow signals indicate the colocalisation of renin with mitochondria. Scale bar: 20 μm. A schematic of the transfected constructs is shown above each image. The leader peptide is depicted in blue, the pro-segment in pink, mature renin in purple and the MTS in red. The red star indicates the insertion of p.L16R mutation. MTS-REN is localised in the mitochondria as the p.L16R renin isoform. (D) Representative western blot analysis showing Mito-YFP when co-expressed with MTS-REN or the p.L16R renin isoform. Images are representative of at least three independent experiments.