MOESM1 of Reduced expression of miR-146a in human bronchial epithelial cells alters neutrophil migration

Additional file 1. Additional material and methods. Figure S1. The expression of miR-146a and selected genes in HBECs from bronchial brushings from patients with asthma. (A) Linear regression analysis between miR-146a expression in asthmatic bronchial brushings (data presented on log2 scale) and eosinophil % among non-epithelial BALF cells. 95% confidence interval (CI) is shown as dotted line. (B) An integrated density value of the signal per area of interest was obtained for 3 areas of bronchial mucosal biopsy samples from 2 non-asthmatic and 3 asthmatic patients. The relative intensity was calculated as percentage and was normalized to the average of control samples (100%). (C) In situ hybridization images of frozen lung tissue biopsy sections from asthmatic patient. Blue color shows the expression of miR-146a, bar = 50 μm. (D) Relative expression of indicated genes from bronchial brush biopsy samples of all included asthmatic patients was measured by RT-qPCR and compared to samples of non- asthmatic controls. Data represent mean ± SEM. Unpaired T-test, *P < 0.05. Figure S2. The expression of miR-146a and selected genes in HBECs transfected with miR-146a or control mimic (24 h) and stimulated for 48 h with TNF-α + IL-17A or left unstimulated (US). Relative miR-146a expression (A) and mRNA (B–D) expression was measured by RT-qPCR. (E) Protein levels of IL-8 and CXCL1 from the supernatants of transfected and stimulated HBECs were measured by ELISA. (F) Neutrophil chemotaxis assay was performed using supernatants of miRNA mimics transfected and TNF-α + IL-17A stimulated HBECs. Migrated neutrophils were counted by flow cytometry. Data represent mean ± SEM. Unpaired t-test, *P < 0.05, **P < 0.01, ***P < 0.001.