Chimeric EGFR ChIP-seq of an MCF-10A clone expressing chimeric EGFR and Erbb2 receptors

Synthetic activation of chimeric EGFR-Erbb2 receptors causes nuclear translocation of the heterodimers at long time scales. To identify genomic loci bound by nuclear EGFR-Erbb2, chromatin from an MCF10A-5E clone coexpressing chimeric EGFR and Erbb2 was prepared with or without synthetic activation and immunoprecipitated for chimeric EGFR by its GluGlu epitope tag. The EGFR-Erbb2 MCF10A-5E clone (B2B1) was grown in overlay culture for six days, stimulated with or without 0.5 µM AP21967 for 24 hours, and prepared for chromatin immunoprecipitation with GluGlu antibody (RRID:AB_2191052) or naïve rabbit IgG as described (Wang et al., Proc. Natl. Acad. Sci. 108:E803-12 [2011]).