Phenotypes of and PI3K-AKT signaling in ptena−/− ptenb−/− medaka embryos.

(A, B) Front (A) and back (B) views of the same pten dko medaka and their littermates at 8 dpf. The genotypes are ptena+/−ptenb−/− (1), ptena+/+ptenb−/− (2), and ptena−/−ptenb−/− (3–5). The eyes of the dko embryos are smaller than those of their littermates, their vasculature is undeveloped with stagnated blood cells being evident (arrows), and their tails are short and curved. (C) Extracts (7.5 μg of protein) prepared from individual medaka embryos of the indicated pten genotypes at 6 dpf were subjected to immunoblot analysis for determination of the pAKT/AKT ratio. Each lane corresponds to an individual. The average pAKT/AKT ratio in dko embryos was about four times that in wild-type embryos (1.41 versus 0.40). Similar results were obtained in other two independent experiments. Given that the embryos shown in (C) were obtained in different experiment from embryos in (A, B), the numbers labeling individual embryos in A and B do not correspond to the numbers shown in C.