Mapping PAK6-AR interacting domain by mammalian two-hybrid assay.

<p>(A) Schematic depiction of full-length PAK6 and boundaries of truncated PAK6mutants encompassing various domains in the left panel. The right panel shows the results of mammalian two-hybrid assays. The interaction was determined by measuring the luciferase activities of HEK 293 cells transiently transfected with PAK6 full-length and deletion mutants fused in-frame downstream of VP16 transactivation domain in a pACT vector and GAL4 DBD fused AR in a pBIND vector. Data represent the mean ± SEM (n = 3). *<i>p</i><0.05, compared to full-length PAK6, one-way ANOVA. (B) The left panel shows the schematic depiction of full-length AR and boundaries of AR truncated mutants encompassing various domains. A mammalian two-hybrid assay was performed by co-transfecting HEK-293cells with pACT-AR or truncated AR mutants, as indicated, with pBIND-PAK6 and pG5-Luc reporter construct. In both sets of experiment, a renilla luciferase reporter activity was used as a transfection internal control. The luciferase activity was normalized against the internal control and the vector basal control groups. Data represent the mean ± SEM (n = 3). *<i>p</i><0.05, compared to full-length AR, one-way ANOVA.