FIG. 3.

GAC63 binds NRs in vitro and in vivo. (A) GST pull-down assays were performed as in Fig. 2A. Full-length ERα and AR were translated in vitro and incubated with GST or GST-GAC63 in the absence or presence of E2 or DHT. (B) COS-7 cells were transfected with 2 μg of pSG5.HA-GAC63 plasmid and 2 μg of pHE0 plasmid (encoding human ERα). Cell extracts were immunoprecipitated with anti-ERα antibody or normal rabbit IgG. The precipitated proteins were detected by immunoblotting with anti-HA antibody. (C) In GST pull-down assays, in vitro-translated ERα 1-281 (containing AF-1 and DBD) or 178-595 (containing DBD and LBD) was incubated with GST-GAC63 in the absence or presence of E2.