Figure 6: DEX and NTHi synergistically induce IRAK-M transcription via inducing the binding of p65 and GR to IRAK-M promoter.
From: Glucocorticoids suppress inflammation via the upregulation of negative regulator IRAK-M
(a–c) Relative luciferase activity (RLA) was measured after the transfection of IRAK-M promoter in pGL3 basic vector and treated with DEX (100 nM) and NTHi in BEAS-2B cells (a,b). (c) MEF cells were transfected with IRAK-M-luc (−500 bp), followed by DEX (1 μM) and NTHi stimulation. (d) Immunoblot shows IRAK-M protein expression in BEAS-2B cells transfected with siRNA p65. (e,f) ChIP assay in BEAS-2B cells treated with DEX and NTHi for 1 h. IRAK-M promoter regions from −38 to +39 bp for GRE, from −231 to −99 bp for κB site were amplified by qPCR. (g) Re-ChIP assay (detailed in Methods). PCR products were detected in agarose gel. (h,i) ChIP assay in BEAS-2B cells treated with IKKβ inhibitor (1 μM), RU486 (1 μM), DEX (100 nM) and NTHi. Data (a–c, n=3; e,f, n=2) are mean±s.d. *P<0.05; t-test.
