Fig. 4
From: Kif17 phosphorylation regulates photoreceptor outer segment turnover
Phospho-mimetic Kif17(S815D) increases disc shedding. a TEM images of 7dpf kif17mw405 larvae that are stably expressing either Kif17-GFP (upper), phospho-mimetic Kif17(S815D)-GFP (middle), or phospho-deficient Kif17(S815A)-GFP (lower) in cones collected at ZT 1.5. Black arrows indicate phagosomes. Scale bar is 2 μm. IS is inner segment. b Quantification of the number of phagosomes for wild-type (n = 5 larvae, 1273 μm of RPE length); kif17mw405 (n = 5, 1170 μm of RPE length); Kif17-GFP, kif17mw405 (n = 10, 2677 μm of RPE length); Kif17(S815D)-GFP, kif17mw405 (n = 10, 2938 μm of RPE length); and Kif17(S815A)-GFP, kif17mw405 (n = 10, 2875 μm of RPE length) larvae. c Quantification of the area of phagosomes for wild-type (n = 5 larvae, 403 phagosomes); kif17mw405 (n = 5, 392 phagosomes); Kif17-GFP, kif17mw405 (n = 10, 530 phagosomes); Kif17(S815D)-GFP, kif17mw405 (n = 10, 1601 phagosomes); and Kif17(S815A)-GFP, kif17mw405 (n = 10, 547 phagosomes) larvae. d Quantification of OS size for wild-type (n = 5 larvae, 268 OS); kif17mw405 (n = 5, 255 OS); Kif17-GFP, kif17mw405 (n = 10, 615 OS); Kif17(S815D)-GFP, kif17mw405 (n = 10, 566 OS); and Kif17(S815A)-GFP, kif17mw405 (n = 10, 605 OS) larvae. e Immunogold labeling of a 7 dpf TaCP:Kif17-GFP zebrafish retina with K62-171c, an antibody against bovine rhodopsin showing a positive labeling of phagosomes (outlined with dashed lines), as also depicted in Additional file 1: Figure S7. Scale bar is 0.5 μm. f Analysis of immunogold labeling of rhodopsin-containing or unlabeled phagosomes in 7 dpf Kif17-GFP, kif17mw405 (n = 5 larvae, 1031 μm of RPE); Kif17(S815D)-GFP, kif17mw405 (n = 5, 1268 μm of RPE); and Kif17(S815A)-GFP, kif17mw405 (n = 5, 889 μm of RPE) larvae. Two-way ANOVA was performed to determine significance in the interaction between the transgene expressed and the concentration of rod or cone phagosomes (p = 0.0264, *) as well as significance in the total number of phagosomes between transgenes (p = 0.0071, **). Bonferroni’s multiple comparisons post-hoc test was performed to determine significance in the differences of unlabeled phagosomes between Kif17-GFP and Kif17(S815D)-GFP (p = 0.0033, **) and between Kif17(S815A)-GFP and Kif17(S815D)-GFP (p = 0.0013, **). There were no differences in numbers of rhodopsin-labeled phagosomes among the transgenes