Figure 2

SKIP increases N- and C-terminal interaction of AR. Mammalian two-hybrid analysis in HEK293 cells that were transiently co-transfected with the GAL4₅E1bTATA-luciferase reporter (250 ng/well), and as indicated, GAL4-DBD-fusion constructs and VP16-fusion constructs of the C- or N-terminus of AR (GAL4DBD-AR 642–917 (125 ng each) and pVP16-AR1-538 (125 ng), respectively) with SKIP-pCGN (100 ng) or empty pCGN (100 ng). Reporter activity was determined in cells with (black bars) and without (grey bars) 1 nM DHT stimulation overnight for 16 hours. Response of cells that did not co-express SKIP (columns 1–4), was markedly lower than that of cells expressing SKIP (columns 5–8). Note the discontinuous y-axis for the relative activity. Data are relative to the control sample (empty vectors without DHT stimulation) as means ± SEM calculated from five independent luciferase reporter experiments, as described in Materials and Methods. Statistically significant difference between indicated means was determined using Student’s t-test.