Fig. 6: IKKα binding to acetyl-His H3 (Lys14) and acting on the promoter region of TSLP in epithelial cells.

a Coimmunoprecipitation of IKKα and Ac-His H3 (Lys14) was performed in cell extracts from 16HBE cells stimulated for 4 h with rhIL-17A (20 ng/ml) in the presence or absence of tiotropium (n = 3). ChiP assay was performed using primers spanning the His H3 binding site of the human TSLP gene promoter: primer 1: 5′–3′; primer 2: 3′–5′; b the cells were stimulated with rhIL-17A (20 ng/ml), alone or in combination, for 4 h. Lane 1: negative control of PCR; lane 2: negative control of immunoprecipitation; lane 3: positive control of PCR; lane 4: untreated cells; lane 5: rhIL-17A (20 ng/ml); lane 6, rhIL-17A + tiotropium (n = 3). c the cells were stimulated with ISs alone or in combination for 4 h. Lane 1: negative control of PCR; lane 2: negative control of immunoprecipitation; lane 3: positive control of PCR; lane 4: untreated cells; lane 5: ISs from COPD patients; lane 6: ISs from COPD patients + tiotropium; line 7: ISs from COPD patients treated with anti-IL-17A Ab. Purified DNA was analyzed by PCR, using control primers specific for the GAPDH promoter. Representative gel images of the experiments are shown