Development of a Novel Molecular Sensor for Imaging Estrogen Receptor-Coactivator Protein-Protein Interactions
Figure 3
Estrogen and anti-estrogen regulation of ERα-AIB1 mediated luciferase fragment complementation.
293 cells were transiently transfected with N-S-AR and EL-S-C (A, C, E, and G) or N-EL and AR-S-C (B, D, F, and H). (A, B) Transfected cells were treated with vehicle (V) or increasing concentrations of E2 for 48 hours prior to quantification of luciferase activities. (C–F) Transfected cells were treated with vehicle (V) or 1 nM E2, in the absence (E2) or presence of increasing concentrations of OHT (C, D) or ICI (E, F) for 48 hours. In all cases data are expressed as firefly luciferase activity normalized to Renilla luciferase activity (± SEM of triplicates). ANOVA was used to determine statistical significance relative to vehicle (A,B) or E2 treatment (C–F; *** p≤0.001, ** p≤0.01, * p≤0.05). (G, H) Lysates from cells treated with vehicle or 1 μM E2, OHT or ICI for 48 hours were immunoblotted using antibodies for ERα, AIB1 or β-actin.