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Loss of <italic>adnp</italic> leads to reduced β-catenin levels and Wnt signaling in zebrafish embryos.a IF staining of FLAG and β-catenin showing the co-localization of FLAG-Adnpa and β-catenin in 3.5 hpf adnpa−/− adnpb−/− embryos injected with mRNAs encoding FLAG-Adnpa. FLAG-Adnpa is predominantly expressed in the nucleus. b IF staining of FLAG and β-catenin showing the co-localization of FLAG-Adnpa and β-catenin in 6 hpf adnpa−/− adnpb−/− embryos injected with mRNAs encoding FLAG-Adnpa. FLAG-Adnpa is predominantly expressed in the nucleus. c Co-IP data showing FLAG-Adnpa interact with β-catenin in 24 hpf adnpa−/− adnpb−/− embryos injected with mRNAs encoding FLAG-Adnpa. d IF staining of β-catenin and HuC showing slightly reduced HuC and β-catenin levels in 16 hpf adnpa−/− adnpb−/− (adnpab−/−) embryos. e IF staining of β-catenin and HuC showing significantly reduced HuC and β-catenin levels in 24 hpf adnpa−/− adnpb−/− embryos. f WB data showing slightly reduced β-catenin levels in 24 hpf adnpa−/− adnpb−/− embryos. g TopFlash luciferase activity assay for total lysates made from 24 hpf control, double mutant embryos and double mutant embryos injected with adnpa mRNAs. Three pools of ten embryos each (n = 3 per group) were lysed with the passive lysis buffer and assayed for luciferase activity. Data are based on two biologically independent experiments, and similar results were obtained. h Representative morphology of 2 dpf adnpa overexpressing embryos. The dorsalized phenotypes (C1–C4) were according to the DV patterning index38. All experiments were repeated at least two times, and similar results were obtained. Shown are representative images. Scale bars: 50 μm in (a, b, d), 100 μm in (e), and 200 μm in (h).
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