Figure 5. The Legless/BCL9 C-terminus binds to core Wnt enhanceosome components.
(A) Top BioID hits showing differential association with wt versus mutant BCL9-BirA* (unweighted spectral counts > 95% probability). (B, C) Western blots of coIPs of wt or mutant GFP-BCL9 with (B) HA-TLE3 or (C) HA-tagged truncations, after co-expression in HEK293T cells (lysed 48 hr after transfection), probed with antibodies as indicated on the left. (D) Top differential BioID hits of B9L-BirA* as in (A). (E) CoIP assays between co-expressed wt or mutant GFP-BCL9, LDB1-FLAG and SSDP-FLAG, as in (B); the band in the top panel corresponds to LDB1-FLAG.
Figure 5—figure supplement 1. HD3-dependent interaction between LDB1 and Legless/B9L.
CoIP assays between co-expressed wt or mutant (A) GFP-B9L or (B) GFP-Lgs, and LDB1-FLAG and SSDP-FLAG, as in main Figure 5E.
Figure 5—figure supplement 2. CRISPR/Cas9-based gene editing strategy for PYGO2 in HEK293T cells.
(A) Cartoon of PYGO2 exon boundaries and targeting site in exon 3; top, sgRNA sequence, with PAM site in bold; underneath, chromosomal location. (B) Western blot of lysates from individual HEK293T clones with PYGO2 KO, probed with antibodies as indicated on the left. (C) CoIP assays between co-expressed wt or mutant GFP-BCL9, and LDB1-FLAG plus SSDP-FLAG in wt or PYGO2 KO cells, confirming that the association of this BCL9 paralog with the ChiLS core complex is partly mediated by its binding to Pygo via HD1.