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Fig. 6 Biochemical activities of zebrafish GADD45β1. Indicated expression constructs (4 μg total: HA-GADD45β1, 2 μg; FLAG-MTK1-N, 2 μg; MTK1, 2 μg; pCS2 vector, 2 μg) were transfected into human 293T cells. Cell lysates were prepared after 24 h. (A) Cell lysates were immunoprecipitated with anti-FLAG gel and analyzed by Western blotting using anti-HA antibody (Top). Whole-cell lysates were used as controls for expression levels. (B) Phosphorylation state of p38 in the transfected cells. Cell lysates were analyzed by Western blotting using antibodies recognizing phospho-p38 and total p38. (C) Live embryos at the 10-somite stage. Synthetic RNA (human MTK1, 300 pg + EGFP, 300 pg) was injected into one blastomere of two-cell-stage embryos, leading to severe somite defects in the injection side.