Fig. 4

shootin1;shootin3 double mutants exhibit reduced number of neuromasts and reduced number of cells in neuromasts. (a) Representative images of wild-type control, shootin1^−/− single mutant, shootin3^−/− single mutant and shootin1^−/−;shootin3^−/−double mutant embryos at 48 hpf. An image of a shootin1^−/−;shootin3^−/− double mutant embryo into which shootin1 and shootin3 mRNAs were injected is also presented at the bottom. Embryos were stained with DAPI. Scale bars: 300 μm. (b) The number of neuromasts obtained from the analyses in (a). The neuromast numbers of wild-type control (n = 15), shootin1^−/−single mutant (n = 19), shootin3^−/− single mutant (n = 16) and shootin1^−/−;shootin3^−/− double mutant (n = 20) embryos were counted at 48 hpf. (c) Rescue experiments of the reduced number of neuromasts in the shootin1;shootin3 double mutants. Injected DKO embryos (DKO + mRNA, n = 9) were observed at 48 hpf. Data for the uninjected wild-type (WT) and DKO mutant embryos in (c) are shared with those in (b). (d) Representative images of the DAPI-stained first deposited neuromasts in wild-type control, shootin1^−/− single mutant, shootin3^−/− single mutant and shootin1^−/−;shootin3^−/− double mutant embryos at 32 hpf. An image of a neuromast in shootin1^−/−;shootin3^−/− double mutant embryo into which shootin1 and shootin3 mRNAs were injected is also shown to the right. Dotted lines indicate the areas of neuromasts, in which small nuclei of neuromast cells cluster. Scale bars: 20 μm. (e) The number of cells in the first deposited neuromasts obtained from the analyses in (d). Wild-type control (n = 14), shootin1^−/− single mutant (n = 6), shootin3^−/− single mutant (n = 11), and shootin1^−/−;shootin3^−/− double mutant (n = 10) embryos were analyzed at 32 hpf. (f) Rescue experiments of the reduced cell number of the double mutant neuromasts. Injected DKO embryos (DKO + mRNA, n = 12) were observed at 32 hpf. Data for the uninjected wild-type (WT) and DKO mutant embryos in (f) are shared with those in (e). Data in (b,c,e and f) represent mean ± SEM; ***P < 0.01; *P < 0.05; ns, nonsignificant.