Interestingly, ESR1 with each of the five LBD alterations identified in this study was inhibited by tamoxifen and fulvestrant in a dose-dependent fashion and did not exhibit the inverted response to anti-estrogens that the synthetic Inv-mut-AA2 mutant did....The IC50 (half-maximal inhibitory concentration) values for both 4-hydroxytamoxifen and fulvestrant were two- to fourfold higher for all mutants compared to wild-type ESR1. Fulvestrant exhibited greater maximal inhibition than 4-hydroxytamoxifen for all the mutants tested (Supplementary Figs. 4 and 5).