Fig. 5

shootin1;shootin3 double mutants exhibit reduced cell number and reduced cell proliferation in the PLLP. (a) Representative images of DAPI-stained PLLP in wild-type control, shootin1^−/− single mutant, shootin3^−/− single mutant and shootin1^−/−;shootin3^−/− double mutant embryos at 32 hpf. An image of a PLLP in shootin1^−/−;shootin3^−/− double mutant embryo into which shootin1 and shootin3 mRNAs were injected is also presented at the bottom. Dotted lines indicate the areas of PLLP, in which small nuclei of PLLP cells cluster. Scale bars: 20 μm. (b) Number of cells in the PLLP of wild-type control (n = 10), shootin1^−/− single mutant (n = 6), shootin3^−/− single mutant (n = 8) and shootin1^−/−;shootin3^−/− double mutant (n = 14) embryos at 32 hpf obtained from the analyses in (a). (c) Rescue experiments of the reduced cell number of double mutant PLLP cells. The injected DKO embryos (DKO + mRNA, n = 7) were analyzed at 32 hpf. Data for the uninjected wild-type (WT) and DKO mutant embryos in (c) are shared with those in (b). (d) Representative images of PLLP stained with EdU and DAPI in wild-type control and shootin1^−/−; shootin3^−/− mutant embryos at 32 hpf. Scale bars: 20 µm. (e) Ratio of EdU-positive cells relative to the total number of cells in the WT (n = 11) and DKO mutant (n = 11) PLLP at 32 hpf obtained from the analyses in (d). Data in (b), (c) and (e) represent mean ± SEM; ***P < 0.01; *P < 0.05; ns, nonsignificant.