Figure 5.

Identification of the elements essential for the progesterone response. Mutations were introduced by site-directed mutagenesis, and sequential deletions were produced by using the restriction sites introduced by site-directed mutagenesis. A, Relative promoter activity, measured by luciferase assays, in mutant (m) and double-mutant (dm) constructs for the different PRE sites. B, Relative promoter activity after the sequential deletion of the −5.7;−4.2 region. C, Relative activity after the mutation of the GATA2 and AP2 sites. In A, B, and C, results were compared with the intact Ednra−5.7;−4.2-luc construct. D, ChIP PCR results from the GATA2 site using the PRB antibody. The lower bands observed in some PCR are the dimers of primers. Lane 1, hPRB; lane 2, hPRB and progesterone; lane 3, as in lane 2 but with an unrelated antibody; lane 4, input control. E, Schematic representation of the sites mutated or deleted in A, B, and C. PRE sites are indicated by their respective number. G, GATA2; A, AP2. #, P < .01; *, P < .05 with ANOVA, comparisons made with control vector.