Fig. 7. Pharmacologic manipulation of TRPV4 modulates mitochondrial transport and the mitochondrial transport protein Miro enhances TRPV4^R269C-mediated toxicity.

a Kymographs of mito-GFP transport in C4da neuron proximal axons in larvae expressing no TRPV4, TRPV4, or TRPV4^R269C treated with 40 nM GSK101 or DMSO. Scale bar, 10 μm. b Fraction of stationary, anterograde-, and retrograde-moving mitochondria from a. Mean ±SEM. For control, TRPV4^WT, and TRPV4^R269C: DMSO n = 9, 10, 10 larvae; GSK101 n = 10, 14, 11 larvae. Two-way ANOVA (p < 0.0001), Tukey’s post hoc test. c Kymographs of mito-GFP transport in C4da neuron axons in larvae expressing no TRPV4 or TRPV4^R269C raised on food containing DMSO or 100 µM GSK219 and treated with 10 µM GSK219 during imaging. Scale bar, 10 μm. d Percentage of stationary, anterograde-, and retrograde-moving mitochondria from c. Mean ± SEM. For control and TRPV4^R269C: DMSO n = 11 and 12 larvae; GSK219 n = 14 and 13. Two-way ANOVA (p < 0.0001), Tukey’s post hoc test. e Percentage ± 95% CI of flies with unexpanded wings when expressing Miro variants ± TRPV4^R269C(low) in N_CCAP. From left to right n = 86, 72, 64, 77, 62, 54, 33, 38, 32, 39, 35, and 41 flies. Χ² test (p < 0.0001) followed by pairwise two-sided Fisher’s exact test. f Representative confocal images of C4da neuron projections in larvae expressing TRPV4^R269C(mod) (as in Supplementary Fig. 2d and e) ± overexpressed Miro variants. Similar results observed in all imaged larvae within each genotype (n = 7, 8, 8, 8, 7, 8, and 8 larvae from left to right) Scale bar, 25 μm. For all panels: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.