Fig. 3.

Inhibition of 11βHSD1-dependent translocation of GR by compound 5. HEK-293 cells were transfected with GFP-GR alone or together with 11βHSD1. Cells were incubated 6 h posttransfection in steroid-free medium for 18 h. Where indicated, cells were preincubated for 30 min with 25 μMGA, 3 or 10 μM of inhibitor 5, followed by incubation for another 40 min with 500 nM cortisone. GFP-GR was detected by immunofluorescence microscopy. By converting inactive cortisone to cortisol, 11βHSD1 mediated nuclear translocation of GR. Inhibition of 11βHSD1 prevented the formation of cortisol and nuclear translocation of GR.