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Fig. 7 Knocking out Axin2, but not Nkd1, increases cytoplasmic levels of β-catenin. (A) Whole cell lysate and cytoplasmic fractions of embryos from wild type and Wnt regulator mutants, with or without 200 pg of wnt8 injected, were collected at 30% epiboly for western blot analysis for cytoplasmic levels of β-catenin. Pan-cadherin was used to confirm purity of the cytoplasmic fraction and Actin was used as a loading control. (B) Because each genotype was run on separate western blots, the increase in cytoplasmic β-catenin when wnt8 was overexpressed was made relative to their uninjected counterpart. Both axin2gh1/gh1 and axin2gh1/gh1;nkd1gh2/gh2 embryos had a greater increase in cytoplasmic β-catenin levels with wnt8 overexpression when compared with wild type embryos. Error bars represent SEM, * = p value < 0.05, one-way ANOVA.