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Fig. S7 Effects of forced expression of constitutively active Ras and Akt. (A) Forced expression of a constitutively active Ras increases Erk1/2 activities in cultured CHO-K1 cells and zebrafish embryos. Western immunoblot analysis results of CHO-K1 cells transfected with HA-RasV12 or zebrafish embryos injected with GFP mRNA or HA-RasV12 mRNA (50 pg/embryo). (B) Expression of constitutively active Ras or Akt at this level does not alter patterning in zebrafish embryos. Expression of emx1 (forebrain), myoD (somite), otx2 (presumptive forebrain and midbrain), pax2a (optic stalk, mid-hindbrain boundary and hindbrain), pax6b (eye field and forebrain) and shh (notochord) in 12-hpf embryos injected with mRNA encoding GFP, HA-Ras V12 or myr-Akt.