Fig. 6.

Dexamethasone Induction of 11β-HSD1 Is Abolished by siRNA against C/EBPβ but Is Unaffected by C/EBPδ siRNA

A, Representative Western blot (20 μg protein/lane) probed with C/EBPβ antibody showing levels of C/EBPβ protein (38-kDa LAP, 20-kDa LIP, and proteolytic products of LAP) 24 h after mock transfection (Mock) or transfection with 80 pmol C/EBPβ siRNA (C/EBPβ) or C/EBPδ siRNA (C/EBPδ) and concurrent vehicle (−) or dexamethasone (+D, 10⁻⁶ m) treatment. The blot was stripped and reprobed with tubulin antibody, as loading control. B, Western blot (20 μg protein/lane) probed with C/EBPδ antibody showing levels of C/EBPδ protein 24 h after mock transfection (Mock) or transfection with C/EBPβ siRNA (C/EBPβ) or with C/EBPδ siRNA (C/EBPδ) and concurrent vehicle (−) or dexamethasone (+D, 10⁻⁶ m) treatment. The blot was stripped and reprobed with tubulin antibody as loading control. C, Real-time PCR measurement of levels of 11β-HSD1 mRNA after mock transfection or transfection with C/EBPβ, C/EBPδ, or scrambled siRNAs. Levels of mRNA in mock-transfected vehicle-treated cells are arbitrarily set to 1. Note logarithmic scale. Data are mean ± sem of at least three independent mRNA samples. ***, P < 0.0001.