Fig. 5

Loss of kif17 diminishes disc shedding. a TEM analysis of phagosome number in 14 dpf wild-type (n = 5 larvae at each timepoint, 1400 ± 97 μm of RPE measured at each timepoint) and kif17^mw405 (n = 5 larvae at each timepoint, 1469 ± 36 μm of RPE measured at each timepoint) zebrafish collected at nine discrete timepoints following light onset (ZT 0). Dark onset begins at ZT 14 (indicated by dark bar). Two-way ANOVA was performed to determine significance in both phagosome number rhythmicity throughout the timepoints (p < 0.0001, ****) as well as between genotypes (p = 0.0011, **). b TEM analysis of phagosome size in 14 dpf wild-type (n = 5 larvae at each timepoint, 93 ± 14 phagosomes measured at each timepoint) and kif17^mw405 (n = 5 larvae at each timepoint, 79 ± 10 phagosomes measured at each timepoint) zebrafish. c Immunogold labeling of rhodopsin-containing phagosomes in 14 dpf wild-type retina at either the morning peak of disc shedding, ZT 1.5 (n = 5 larvae, 1738 μm of RPE), or the evening peak, ZT 15.5 (n = 5, 1341 μm of RPE). Two-way ANOVA was performed to determine significance in the interaction between the time of day and the concentration of rod or cone phagosomes (p = 0.0082, **). There were no statistical differences in the total number of phagosomes between the morning or night peak. Bonferroni’s multiple comparisons post-hoc test was performed to determine significance in the increased of unlabeled phagosomes between morning and night (p = 0.0158, *). d TEM analysis of phagosome number in 2–4 month wild-type (n = 3 mice at each timepoint, 979 ± 42 μm of RPE measured at each timepoint) and Kif17^tm1b(Bjc) (n = 3 mice at each timepoint, 940 ± 45 μm of RPE measured at each timepoint) mice collected at eight discrete timepoints following light onset (ZT 0). Dark onset begins at ZT 12 (indicated by dark bar). Two-way ANOVA was performed to determine significance in both phagosome number rhythmicity throughout the timepoints (p = 0.0300, *) as well as between genotypes (p = 0.0096, **). e TEM analysis of phagosome size in 2–4 month wild-type (n = 3 mice at each timepoint, 54 ± 7 phagosomes measured at each timepoint) and Kif17^tm1b(Bjc) (n = 3 mice at each timepoint, 35 ± 4 phagosomes measured at each timepoint) mice. f Immunogold labeling of rhodopsin-containing phagosomes with B630 rhodopsin antibody in mouse retina at either the morning peak of disc shedding, ZT 1.5 (n = 5 mice, 554 μm of RPE), or the evening peak, ZT 13.5 (n = 5, 594 μm of RPE). Two-way ANOVA was performed to determine no significance differences in the interaction between the time of day and the concentration of rod or cone phagosomes (p = 0.5700) or in the differences in phagosome number between the morning or night peak (p = 0.8076), but there was a significant increase in the total number of rod phagosomes compared to cone phagosomes (p = 0.0074, **)