Fig. S5

Rap1b Regulates PLPM Cell Migration, Related to Figure 4. (A) Left: Expression patterns of desma in sibling (WT, upper) and rap1bncv124 (lower) embryos at 14 hpf. Dorsal views with anterior to the top. Right: Average number of somites in sibling (upper) and rap1bncv124 (lower) embryos. The number of somites were counted based on the expression of desma. WT, n=31; rap1bncv124, n=28. ss, somite stage. (B) Confocal fluorescence images of 16 hpf Tg(fli1a:GFP) embryos. Dorsal views with anterior to the upper-left. Left, merged image of DIC and GFP; right, GFP image. Arrows indicate somite boundaries. Note that medial PLPM cells migrate along somite boundaries. (C) Number of fli1a:GFP-positive PLPM cells or cell clusters left behind group of migrating cells in wild type (blue circles) or rap1bncv124 (pink squares) embryos at the 11 indicated developmental stages, as observed in Figure 4A. Data are means ± s.d. WT, n=11; rap1bncv124, n=18. *, p < 0.05; **, p < 0.01. (D) Time-lapse confocal images of PLPM cell migration in sibling (WT) and rap1bncv124 Tg(fli1a:GFP) embryos injected without (Uninject) and with dld mRNA from 14-19 hpf. Dorsal views with anterior to the top. Arrowheads indicate the cells that were left behind the group of migrating cells. (E) Number of fli1a:GFP-positive PLPM cells or cell clusters left behind group of migrating cells in wild type embryos injected without (blue circles) or with (yellow triangles) dld mRNA and rap1bncv124 embryos injected without (purple squares) or with (grey triangles) dld mRNA at the indicated developmental stages, as observed in (D). WT, n=7; rap1bncv124, n=9; WT+dld mRNA, n=6; rap1bncv124+dld mRNA, n=8. Scale bars: 200 μm (A); 100 μm (B, D).