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Fig. 3 stap2b knockout embryo phenocopy morphant's phenotype. (A) Schematic showing the Cas9-gRNA targeted site in stap2b. The protospacer-adjacent motif (PAM) sequence is GGG, and the targeted sequence is labeled in red. (B?D) Bright field images of wild-type (WT), stap2bATG morpholino injected embryos (stap2bATG MO) and stap2b-knockout embryos (stap2b?3). (E?G) At 30 hpf, compared to wt controls, knockout of stap2b (stap2b?3) showed an ISV growth defect similar to the phenotype of stap2b morphants, which was induced by morpholino injection (hollow arrowhead in C and D). (n = 12 in WT embryos, n = 13 in stap2b morphant embryos, and n = 14 in stap2b knockout embryos). (E??G?) Loss of stap2b in morpholino and stap2b-knockout Tg (kdrl:mCherry; fli1a:negfp y7) embryos showed that the cell number per ISV was less than that in WT embryos. (H?J) Meanwhile, CVP loop formation was lower (arrows in I and J) in stap2b morphants and stap2b?3 mutants than in WT control (H). (K) Quantification of the percentage of completed ISVs in stap2b morphant and stap2b-knockout embryos was 55% and 35% reduced compared to WT embryos. (L and M) Quantification of the average cells per ISV and CVP structure in WT embryos, stap2b morphants and stap2b-knockout embryos. Scale bars are 200 ?m for B-D and 100 ?m for E?G, E??G? and H-J. (***Refers to p < .001 and **refers to p < .01)