FIGURE 1.
Coimmunoprecipitation of FKBP51 with both GRα and -β and of FKBP52 with only GRα in NTM-5 cells. NTM-5 cells were cultured in 10% FBS-DMEM to complete confluence, shifted to serum-free DMEM, and treated with vehicle (ethanol) or 100 nM DEX for 2 hours. The cytoplasmic (Cyto) and nuclear (Nuc) fractions were isolated. Anti-FKBP51 or anti-FKBP52 immunoprecipitates of cytoplasmic (100 µg) and nuclear (50 µg) proteins were subjected to electrophoresis in SDS-polyacrylamide gradient gels followed by Western immunoblot analysis with anti-GRα or anti-GRβ antibodies. (A) Coimmunoprecipitation with anti-FKBP51 or anti-FKBP52 antibodies, and Western immunoblot analysis with a specific anti-GRβ antibody. GRβ was detected as a 90-kDa protein band. (B) Coimmunoprecipitation with anti-FKBP51 or anti-FKBP52 antibodies and Western immunoblot analysis with a polyclonal GRα antibody. GRα was detected as a 95-kDa protein band. The experiment was performed independently three or more times.