Fig. 1

MCU expression is limited in the retina, particularly in cones. a Western blot of zebrafish tissue lysate, enriched for mitochondrial proteins and probed for MCU, mitochondrial cytochrome oxidase (MTCO1), and succinate dehydrogenase (SDH). Samples were pooled from either four retinas, two hearts, or one brain and performed with n = 3 replicates of each pool. Each lane contains 8 µg of protein. The lower panel shows quantification of the replicates, normalized to retina tissue. The mean is reported and bars = standard error. **p < 0.01, ***p < 0.001 using ANOVA followed by Dunnett post hoc test (comparison with retina). b Within each lane from gels analyzed in panel A, the ratios of MCU signal to the mitochondrial proteins MTCO1 and SDH were determined. Values were normalized relative to retina tissue. The mean is reported and bars = standard error. *p < 0.05, **p < 0.01 using ANOVA followed by Dunnett post hoc test (comparison with retina). c qRT-PCR quantification of relative mRNA of MICU proteins (relative to reference gene Ef1α and/or b2m, see “Methods”) across retina, heart, and brain tissues. The mean is reported and bars = standard error. *p < 0.05, **p < 0.01, ***p < 0.001 and ns = not significant using ANOVA followed by Dunnett post hoc test (comparison with retina). d Mitochondria-enriched retinal lysate of WT and pde6c^−/− cone deficient retinas. Each lane is from lysate of two pooled retinas from a single fish. The right panel shows quantification of replicates; n = 4 fish. Each lane contains 30 µg of protein. The mean is reported and bars = standard error. Ns = not significant using Welch’s t-test. e Relative quantification of the ratio of MCU to mitochondrial proteins MTCO1 and SDH in WT and pde6c^−/− cone deficient retinas from experiments shown in D. The mean is reported and bars = standard error. *p < 0.05, **p < 0.01 using Welch’s t-test