Supplementary Figure 6: dzip1l zebrafish morphants display ciliopathy-associated phenotypes.

(a) Wholemount of a 20 hpf zebrafish embryo showing dzip1l expression in the nasal placode (short arrow) and pronephric duct (long arrow). (b) dzip1l expression in MCC precursors in the pronephric duct at 24 hpf (arrows). Inset shows expression in individual MCC precursors. (c) dzip1l expression in the nasal placode at 24 hpf (arrows). (d) Wild-type control embryo at 3 dpf. (e) Two otoliths in the otic vesicle of a wild-type embryo at 3 dpf. (f) dzip1l translation blocking morpholino (dzip1l MO) injected embryo at 3 dpf with shortened, slightly curved body, hydrocephalus (arrow) and reduced retinal size (arrowhead) (g) Three otoliths in the otic vesicle of a dzip1l morpholino (i4e5) injected embryo at 3 dpf. (h) Wild-type glomeruli (upper left panel) and cystic dilated glomeruli in dzip1l MO injected embryos (lower left panel, white asterisk) at 48 hpf. (i) Incidence of glomerular cysts and otolith deposition defects in control (co MO), translation blocking (TB MO) and splice site (i1e2 MO and i4e5 MO) morpholino injected Tg(wt1b::GFP) embryos at 48 hpf (for glomerular analysis n = 425 embryos for translation blocking morpholino, n = 349 embryos (i1e2) and n = 384 embryos (i4e5) for splice morpholinos; t(13)=5.68, ***p=0.0008 (i1e2 MO), t(17)=8.36 and t(14)=8.27, ****p <0.0001 (TB and i4e5 MO); for otolith analysis n = 243 embryos for translation blocking morpholino, n = 236 embryos (i1e2) and n = 326 embryos (i4e5) for splice morpholinos; t(7)=5.42, **p=0.0011 (TB MO), t(6)=14.93, ****p<0.0001(i1e2 MO), t(7)=4.34, **p=0.003 (i4e5MO). Statistical analysis based on an unpaired two-tailed Student`s t-test. Error bars, s.e.m. (j) Co-injection of zebrafish dzip1l mRNA (ddzip1l) can rescue dzip1l morpholino caused phenotypes; n = 463 embryos for morpholino and n = 313 embryos for mRNA co-injected embryos; t(9)=9.06, ****p<0.0001. Scale bars in a-g = 100μm; h = 50μm; each data point represents a single experiment.