ChIP-qPCR Protocol for Lef1 Binding

ChIP-qPCR was performed as described previously^{5 (link)}. Cells were crosslinked with formaldehyde and the reaction was quenched with glycine. Cells were lysed and chromatin was digested with HindIII restriction enzyme (Thermo Fisher FD0505). ChIP grade anti-Lef1 mouse antibody (Merck 17–604) was coupled with Dynabeads coated with sheep anti-mouse antibody (ThermoFisher 11201D). Beads were added to chromatin and bound to DNA associated Lef1. After this immunoprecipitation (IP) step, precipitated chromatin was treated with Proteinase K (Thermo Fisher 4,333,793). DNA was isolated using sodium acetate method. PCR primers were obtained either from the Epitect array (Qiagen 334,211) or as reported in the SI Appendix Table 1.