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. 2012 Aug 13;287(41):34809–34824. doi: 10.1074/jbc.M112.372797

FIGURE 5.

FIGURE 5.

Requirement for MAGE-11 in PR-B up-regulation of FKBP5 but not RASD1. A, top, IKPRB cells were treated in charcoal-stripped serum-containing medium with 20 nm R5020 for 0, 8, and 24 h. Cell extracts (40 μg of protein for FKBP5 and β-actin, 0.1 mg protein for PR-B) were probed using FKBP5, PR, and β-actin antibodies. Bottom, IKPRB cell extracts (0.1 mg of protein/lane; lane 1) and LAPC-4 cell extracts (40 μg protein/lane; lane 2) prepared in immunoblot lysis buffer were probed on transblots using PR sc-7208 (1:250 dilution) and FLAG-MAGE antibody-1 (10 μg/ml) plus MAGE-11-(94–108) antibody (5 μg/ml). IKPRB cells (B, C, and E) and IKPRAB cells (D and F) were selected using puromycin for lentivirus expression of MAGE-11 shRNA-827, -947, and -964, nonspecific (NS) shRNA, or no virus addition (−). mRNA was assayed by quantitative real-time RT-PCR as described under “Experimental Procedures.” MAGE-11 mRNA was assayed in IKPRB cells incubated without progesterone (B). FKBP5 (C) and RASD1 mRNA (E) was assayed in IKPRB cells incubated for 6 h with and without 10 nm progesterone. FKBP5 (D) and RASD1 mRNA (F) was assayed in IKPRAB cells incubated for 12 h with and without 10 nm progesterone. Error bars, S.E.