ER alpha coregulator peptide screening profiling by MARCoNI (Microarray Assay for Real-time Coregulator-Nuclear receptor Interaction) assay from human embryonic kidney HEK293 cells after tranfected with ER alpha or Q375 mutations

Estrogen insensitivity syndrome (EIS) arises from rare mutations in ERα resulting in the inability of estrogen to exert its biological effects. Due to the rarity, mutations in ESR1 gene and the underlying molecular mechanisms of EIS have not been thoroughly studied. We used PamChip array with 154 coregulator motifs (PamGene #88101) array to compare the peptide profiles between the WT and 5 of Q375 mutants in HEK293 cells after transferred with ER alpha or Q375 mutatnts. 93 ligand-modulated peptides were reported here. HEK293 cells after tranfetced with WT ER or Q375 mutants were collected for cell lysate. cell lysate of ERα genotype expressing cells was incubated with or without ligand on a PamChip array with 154 coregulator motifs (PamGene #88101). Ligands: veh (DMSO, control), E2 (17-b-Estradiol), CME (11b-chloromethyl estradiol), Et-E2 (11b-ethyl estradiol), THC (diethyl-tetrahydrochrysene), DES (diethylstilbestrol) or Hex (meso-hexestrol).