Fig. S3

Phenotypes of the ninl atgMO.

(a) Representative clutch of zebrafish larvae at 2dpf injected with the phenotypic dose of ninl atgMO (2ng/nl). (b) Titration curve for the ninl atgMO illustrating the distribution of phenotypes in 2dpf larvae at two different concentrations: at 1ng/nl, a minority of injected larvae present a curved body shape (10%) and/or ventriculomegaly (20%) (n = 19). At ~2ng/nl, on average 66% of injected larvae present a curved body shape and ~40% present ventriculomegaly and/or pronephric cysts (n = 84). 95% Confidence Interval bars are shown. (c) Representative 2dpf-old ninl atg-morphant displaying curved body shape, ventriculomegaly and pronephric cyst (arrow). (d-e) Dorsal view of 2dpf larvae showing the normal morphology of the brain folds in wild-type (d) and the enlarged ventricle in morphants (e). (f-g) Transgenic Tg(wt1b:EGFP) zebrafish line used to highlight the larval pronephros, shows the morphology of the fused glomerulus and proximal tubules in wild-type (f) and the dilatation of the region in ninl morphants (“kidney cysts”, white arrow in g). (h) A clutch of ninl morphants at 4dpf. (i-j) TUNEL assay on 4dpf cryosections through retinas from ninl morphants shows the range of cell death detected (curved larvae in (h) were sectioned for the TUNEL assay). Note the absence of TUNEL-positive cells in the photoreceptor (PR) cell layer in the morphants. (k) The ift88-/- retina is used as a positive control, given the known death of photoreceptors in this mutant at 4dpf. (l) Cryosection through a 4dpf brain in a morphant larva displaying a dilated brain ventricle (v) shows no significant neuronal cell death. Scale bars represent 500 μm in (a) and (h), 100 μm in (c-g), 10 μm in (i-k) and 30 μm in (l). PR PhotoReceptors, INL Inner Nuclear Layer.