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Fig. 9 a Overview and schematic diagram of human PRKCD, zebrafish prkcda and prkcdb proteins. Percentage identity of respective domains on comparison with human counterpart shown below the zebrafish domains. Also shown is the percentage identity of the orthologues amongst each other. b Dorsal view of spatial expression pattern of prkcda and prkcdb at 5 dpf as demonstrated by whole-mount in situ hybridisation using a 5’UTR probe. Scale bar = 200 μm. c Representative confocal images of cryosections taken from transgenic tandem-tagged mitofish at 3 dpf, fixed and stained for nuclei (Hoechst) and endogenous Prkcd. The image shows the section of a larval zebrafish hindbrain. Representative ×40 image of the zebrafish hindbrain section was taken on Zeiss LSM 800, scale bar = 20 µm. d Representative ×20 confocal images (Zeiss LSM 800) of cryosections taken from transgenic tandem-tagged mitofish at 3 dpf, fixed and stained for chondroitin sulphate. The markings (white dashed line) represent the hindbrain region, following the chondroitin staining. Scale bar = 50 µm. e Representative immunoblots of Prkcd and β-actin on whole embryo lysates from wild-type and single or double prkcda/prkcdb KO (DKO) animals. f Representative ×20 and ×60 confocal images of cryosections taken from control (guide only) and prkcd_ab DKO transgenic tandem-tagged mitofish larvae treated with DMSO or with DMOG at 3 dpf for 24 h. Images are from within the hindbrain region of the respective larvae as marked (dashed white line). Scale bars = 50 μm. g Quantitation of the number of red puncta from a 50-μm hindbrain region (as marked in f) from control and prkcd_ab DKO tandem-tagged mitofish larvae. Plots demonstrate data distribution and median value (red line). Significance was determined by two-way ANOVA followed by Tukey’s post-test to compare all groups. h Motility analysis of zebrafish embryos at 5 dpf using the “Zebrabox” automated videotracker (Viewpoint). The assay was carried out during daytime and consisted of one cycle of 30 min exposure to light followed by 30 min of darkness. Data represent mean distance moved ± SEM. Each group consisted of 43–124 larvae from n = 9 independent experiments. Significance was determined by two-way ANOVA followed by Dunnett’s post-test to the control (WT). Significance are denoted where *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, n.s. = not significant. For precise P values, see the source data file.