Figure 5.

Fis1 ∆SKY variants loss of function is rescued by TBC1D15 expression. The impact of YFP-TBC1D15 expression on mitochondrial morphology and DRP1 localization was determined from experiments shown in Figures 3 and 4; HCT116 cells co-overexpressing FIS1 with either mitoYFP (-YFP-TBC1D15, from Fig. 3 experiments), or YFP-TBC1D15 (+YFP-TBC1D15, from Fig. 4 experiments) were analyzed for mitochondrial morphology and DRP1 localization. A, representative confocal images showing merged anti-DRP1 (magenta) and anti-TOM20 (yellow) from single channel images before (left panel) and after (right panel) transfection with YFP-TBC1D15. Note for ΔSKYD49G, the Figure 3C image are reused in A. The scale bar represents 10 μm (magnified inset scale bar represents 5 μm) with fluorescence intensities adjusted for clarity. B, violin plots of average mitochondrial component area in absence (left panel, from Fig. 3B) and presence (right panel) of YFP-TBC1D15 coexpression. C, violin plot of the colocalization between TOM20 and DRP1 from single cell maximum intensity projections was measured using Pearson’s correlation coefficient area in absence (left panel, from Fig. 3D) and presence (right panel) of YFP-TBC1D15 coexpression. D, correlation plot to determine the relationship between mitochondrial component area and mitochondrial DRP1 in absence (left panel, from Fig. 3E) and presence (right panel) of YFP-TBC1D15 coexpression. Each point in B and C represents a single cell and each circle in D represents the population means and are colored based on the FIS1 expression levels determined from mean fluorescence intensity per cell. Data represent three biological replicates with p values calculated from two-way ANOVA analyses, followed by TUKEY honest significant differences (HSD). FIS1, fission protein 1; HCT, human colorectal carcinoma.