Fig. 4

HOMER3 functions as a scaffold platform to facilitate c-Src and β-Catenin interaction. a The interaction between HOMER3 and β-Catenin was analyzed by IP assays. b Interactions between β-Catenin and truncated HOMER3 proteins. c Up panel: Schematic illustration of β-Catenin truncated constructs. Low panel: 293 T cells were transfected with indicated Flag-β-Catenin truncations and HA-HOMER3, followed by IP assays with Flag-beads to examine their interaction with HOMER3. d Reciprocal IP assays of c-Src and β-Catenin in MDA-MB-231 cells with or without HOMER3 silencing. e In vitro protein binding assays with purified HOMER3, β-Catenin and c-Src protein. The purities of HOMER3, c-Src and β-Catenin were examined by SDS-PAGE and Coomassie Blue Staining. f IP assays of c-Src in HOMER3 silencing MDA-MB-231 cells with restoration of full-length or truncated HOMER3 constructs. g Tyr phosphorylation of β-Catenin in HOMER3 silencing cells with restoration of full-length or truncated HOMER3 constructs. g Normalized luciferase activities of specific TOP-Flash over non-specific FOP-Flash relative renilla luciferase units (RLU) in indicated cells. h Fluorescence immunostaining of β-Catenin expression in HOMER3 silencing MDA-MB-231 cells with restoration of indicated HOMER3 constructs