Fig. 5

Effect of RAD001 (RAD), neratinib (Ner), or their combination on oestrogen receptor (ER)-mediated transactivation and recruitment of the ER basal transcription machinery. a Cell lines were co-transfected with EREIItkLuc and pCH110 and treated for 24 h with RAD001 and neratinib in the absence of E2 (DCC). Luciferase activity was normalized by β-galactosidase from triplicate wells and fold-changes expressed relative to the DCC control. b ChIP analysis to determine the effect of neratinib, RAD001, or the combination on recruitment of ER to the TFF1 promoter in wt-HCC1428 and HCC1428-LTED. Error bars represent mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001. Concentration used for transactivation assay and ChIP: wt-MCF7 (2 nM RAD001, 500 nM neratinib); MCF7-LTED (4 nM RAD001, 650 nM neratinib); wt-SUM44 (3 nM RAD001, 700 nM neratinib); SUM44-LTED (3 nM RAD001, 700 nM neratinib); wt-HCC1428 (3 nM RAD001, 700 nM neratinib); HCC1428-LTED (10 nM RAD001, 300 nM neratinib). ns not significant