Fig. 5. Expression of dnPI3K in PGCs results in their localization to ectopic positions. (A,B) Expression of dnPI3K-nos1-3'UTR results in PGCs in ectopic positions at 24 hpf (arrowheads in B) as compared with control embryos where PGCs are located in their proper positions (arrow in A). Embryos were co-injected with GFP-nos1-3'UTR to label the PGCs. (C,D) dnPI3K-nos1-3'UTR-injected embryos show normal morphology (D) as demonstrated by in situ hybridizations of embryos at the 6-7-somite stage stained with sdf-1a (red) and nos1 (blue) and compared with a control embryo (C). High-magnification inset in C shows PGCs in regions of sdf-1a expression in control embryos whereas inset in D shows a subset of PGCs in a region of non-sdf-1a expression in dnPI3K-injected embryos. Arrows in B,D and high-magnification inset in F indicate the presence of PGCs in their proper position in dnPI3K-injected embryos. (E,F) PGCs located in ectopic positions in dnPI3K-injected embryos show normal expression of nos1 mRNA (arrowheads in F) similar to PGCs in control embryos (arrow in E). (G) Graph illustrating the percentages of embryos with ectopic PGCs. 2% of control embryos (green bar) had 2 PGCs located in ectopic locations at 24 hpf as compared with 63% of dnPI3K-nos1-3'UTR injected embryos (red bar) showing 2 ectopic PGCs by 24 hpf. (H) Graph illustrating the average speed of PGC migration in control embryos (green bar) and dnPI3K-nos1-3'UTR-injected embryos (red bar). Control PGCs migrate at an average speed of 1.7 µm/min (n=23) compared with PGCs that express dnPI3K, which migrate at a rate of 1.3 µm/min (n=21; P<0.05).