Figure 4.

Mutational analysis of two potential PREs in the first intron of FKBP5. A, Computer analysis of the mouse FKBP5 gene predicted two potential PR-binding sites, PRE1, located at +1069 to +1087 bp, and PRE2, located at +1122 to +1138 bp, as well as a GATA site (in box) in the first intron. B, A series of constructs (M1–M8) were generated by mutating the wild-type murine FKBP5 intronic sequences at the positions depicted (see Materials and Methods for details) and were transfected into HeLa cells along with PR. After 24 h of R5020 treatment, cells were harvested and lysed, and whole-cell extracts were assayed for luciferase and β-galactosidase activity. The values were normalized for β-galactosidase and are expressed as fold luciferase activity with respect to vehicle-treated cells. Data shown here represent the mean ± se of three independent experiments performed in duplicate. Fold inductions are denoted over each bar. WT, Wild type.