Fig. 4: The affinity of indicated compounds to the AR and their regulation of AR transcriptional activity. | Cell Death & Disease

Fig. 4: The affinity of indicated compounds to the AR and their regulation of AR transcriptional activity.

From: The beneficial androgenic action of steroidal aromatase inactivators in estrogen-dependent breast cancer after failure of nonsteroidal drugs

Fig. 4

a AR fluorescence polarization-based competitive hormone-binding assays. Rat AR ligand-binding domains tagged with a His glutathione S-transferase epitope (His-GST-ARLBD) were used at final concentrations of 25 nM. The fluorescently labeled AR ligand, Fluormone AL Red, was used at a final concentration of 2 nM. The competing test compounds were DHT, 4-OHA, 4-OHT, 17-HEXE, EXE, and Mibo as indicated. Point, mean of triplicate determinations; bars, 95% confidence intervals. Curve fitting was done using GraphPad Prism software (version 7.00). IC50s corresponding to a half-maximal shift in polarization values of the test compounds were determined using the maximum and minimum polarization values of the DHT-competitive-binding curve for AR. b Tested compounds regulate AR transcriptional activity at a concentration of 100 nM. ARE(3×)-regulated dual-luciferase activity in U2-OS cells under steroid-free conditions. Cells were transiently transfected with pcDNA3.1-AR and ARE3-luc2P (firefly luciferase reporter plasmids) and the internal normalization control pRL-TK (Renilla luciferase reporter plasmid). Six hours after transfection, cells were treated as indicated. Then they were assayed 48 h after transfection for dual-luciferase activity. Data shown are the mean of triplicate determinations and associated SEM; bars, SEM. *P ≤ 0.05, **P ≤ 0.01 vs. control

Back to article page