FIGURE 10.

Dependence on PR-B NH₂-terminal ¹¹⁰LLXXVLXXLL¹¹⁹ motif for down-regulation and transcriptional activation by MAGE-11. A, human PR-B NH₂-terminal 164 amino acid residues contain activation function 3 (AF-3) and an ¹¹⁰LLXXVLXXLL¹¹⁹ motif not present in PR-A. PR-A and -B have an identical DNA-binding domain (DBD) and ligand-binding domain (LBD). B, p5M-PR-B and L110A,L111A, V114A,L115A, and L118A,L119A mutants and p5M-PR-A (6 μg) were expressed in COS cells with and without 10 nm progesterone. The transblot of cell extracts (30 μg of protein/lane) was probed using PR sc-7208 antibody. C, p5M-PR-B wild type (WT) and L110A,L111A, V114A,L115A, and L118A,L119A mutants (0.5 μg) were expressed in COS cells with and without 1 μg of pSG5-MAGE in the absence of progesterone. The transblot of cell extracts (30 μg protein/lane) was probed using PR sc-7208 (1:250 dilution) and FLAG-MAGE-1 antibodies (0.5 μg/ml). D and E, PR-B WT; L110A,L111A, V114A,L115A and L118A,L119A mutants; and PR-A (25 ng) were expressed in CV1 cells with 3 μg of pIE2-Luc FKBP5 luciferase reporter vector and 0.5 μg of pSG5 empty vector (−) or 0.5 μg of pSG5-MAGE (M) (D) or 0.5 μg of pSG5-HA-p300 (P) (E). Cells were incubated with and without 1 nm progesterone. Error bars, S.E.