Figure 3.

FKBP52 potentiation of interhelical proline mutants within the GR LBD. A–F, Yeast were transformed with a β-galactosidase reporter pUCΔSS-26x, a receptor expression plasmid for either wild-type (wt) or mutated rGR and an FKBP52 expression plasmid or empty vector control. Aliquots of yeast culture were dosed with 25 nM (wt) or 5 μM (all mutants) of DOC. The reporter expression of each receptor in the presence or absence of FKBP52 is presented. Graphs are representative of 3 independent experiments. G, Levels of transformed protein were determined by running yeast protein extracts through a 10% SDS-PAGE gel and a Western analysis performed by transferring overnight to a nitrocellulose membrane and probing sequentially for GR (H-300 antibody), FKBP52 (Hi52c antibody), and the internal control L3 (α-L3 antibody). Vertical lines separate membranes derived from different gels. Abbreviation: RLU, relative light units.