BBS4 is required for normal zebrafish development and rhodopsin localization. A: Representative examples of whole-mounted wild-type (WT) embryos, morpholino-injected (MO) embryos, embryos injected with wild-type (wt) or mutant (mut) human BBS4 mRNA, or embryos coinjected with morpholino and mRNA. Dorsal view (top row) and lateral view (bottom row) are shown of embryos at the 12–14 somite stage following in situ hybridization with pax2a and myoD riboprobes. Embryos were categorized phenotypically based on shortened body axis (anterior and posterior ends marked by red triangles) and notochord defects (red arrow). B: Quantification of the efficiency of rescue from gastrulation defects following coinjection of BBS4 morpholino (MO) and mRNA. The number of animals analyzed for each group is noted above each bar. C: Retinal cryosections of 5 dpf zebrafish retinas stained for rhodopsin (red). White arrows indicate rhodopsin mislocalization (Scale bar=10 μm).

E235Q mutant protein is produced and correctly localizes to the pericentriolar region in HeLa cell culture. Immunofluorescence of FLAG-tagged wild-type and mutant BBS4 protein in HeLa cells. Cells were stained using an anti-FLAG antibody (green) for BBS4 expression and anti-γ tubulin antibody (red) for centrosomal localization. Nuclei were stained with DAPI.