Journal of Toxicology / 2009 / Article / Fig 4

Research Article

Molecular and Biochemical Effects of a Kola Nut Extract on Androgen Receptor-Mediated Pathways

Figure 4

Biz-2 competes for AR binding in LNCaP cells. AR protein from LNCaP cells, cells transfected with pCMV-AR, or purified recombinant AR protein was incubated with 2 nM [3H] R1881 with or without increasing concentration of Biz-2 for 18 hours at 4°C. Bound and free [3H] R1881 were separated using the HAP assay (see Section 2). Each data point represents the mean ± SEM of a representative experiment performed in triplicate. The IC50 was calculated from the competition curve using a one-site competition model. was calculated by = IC50/(1 + [ligand]/ ) in which and ligand concentration [ligand] were set to 1 and 2 nM, respectively. (a) Single-point assay in the presence of 500X excess R1881 (number above the bar is % of total binding). (b) Displacement curve of [3H] R1881 by Biz-2 (insert is displacement curve of [3H] R1881 by inert R1881). N = 2 separate experiments performed in triplicate.