381824.fig.002a
(a)
381824.fig.002b
(b)
381824.fig.002c
(c)
Figure 2: Nuclear localization and transactivation ability of N-TAP-mAR. (a) Immunostaining of a stable mesenchymal cell line M7 and a stable epithelial cell line P17 expressing N-TAP-mAR. The FLAG antibody detects tagged androgen receptor only. N20 antibody recognises both endogenous AR and N-TAP-mAR. Nuclei staining with DAPI indicates the percentage of cells expressing NTAP-mAR. (b) The ability of the modified N-TAP-mAR and WT-AR to activate a glucocorticoid response element (GRE) was assayed in COS-1 cells in a transient transfection luciferase assay. GRE promoter activation, shown here on the -axis as luciferase/renilla ratio, was similar for the tagged and nontagged androgen receptor. (c) Tagged androgen receptor expression (N-TAP-mAR) was elevated in the Western blot done with protein extracts from the transactivation samples relative to WT.