ADNP stabilizes β-catenin during ESC neural differentiation.a WB showing total β-catenin levels in day 3 control and Adnp−/− ESC-derived neurospheres. b WB showing β-catenin levels in the cytoplasmic and the nuclear fraction of lysates from day 3 control and Adnp−/− ESC-derived neurospheres. Note that β-catenin levels were significantly reduced in the cytoplasmic but slightly reduced in the nuclear fraction of lysates. c Time course WB assay showing total and phosphorylated β-catenin levels during control and mutant ESC differentiation toward a neural fate. d Representative image of IF staining of β-catenin in day 19 ESC-derived neuronal cultures; e restoring FLAG-ADNP in mutant ESCs could rescue the β-catenin levels in day 6 neurospheres. f Representative WB showing β-catenin levels in the combination treatment of CHIR and MG132. g Representative WB showing the ubiquitylation levels of β-catenin in control and Adnp−/− ESCs. h Representative WB showing that total β-catenin levels were slightly elevated and p-β-catenin levels were slightly reduced in day 3 neurospheres from FLAG-ADNP overexpressing ESCs. i Representative WB showing the expression of the indicated proteins in HEK293T cells that were transfected with an increasing dose of ADNP. WB and IF staining experiments have been repeated at least two times and similar results were obtained.
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