Additional file 2: of The anti-proliferative and anti-inflammatory response of COPD airway smooth muscle cells to hydrogen sulfide

Figure S2. Further examples of the effect of the H2S donor, NaSH on CBS and MPST protein expression, and activation of extracellular signal–regulated kinase (ERK)–1/2 and p38 mitogen-activated protein kinase (MAPK) in human ASM cells from non-smokers, smokers and COPD patients. ASM cells were incubated with FCS (2.5%) for 1 h and NaSH (100 μM) was added for another 24 h. CBS, MPST (A), Total and phospho–ERK-1/2, total and phospho-p38 and β-actin (B) were detected by Western blotting. (JPG 212 kb

Additional file 1: of The anti-proliferative and anti-inflammatory response of COPD airway smooth muscle cells to hydrogen sulfide

Figure S1. Immunohistochemistry staining of CSE, CBS and MPST in bronchial biopsies from non-smokers, smokers and COPD patients. Photomicrographs showing representative photomicrographs of cystathionine-γ-lysase (CSE), cystathionine-β-synthase (CBS) and 3-mercaptopyruvate sulphur transferase (MPST) staining in the bronchial mucosa from control non-smokers, control smokers with normal lung function and mild/moderate COPD patients. Immune-stained airway smooth muscle cells are indicated by brown staining. Results are representative of those from 13 non-smokers, 14 smokers with normal lung function, 15 mild/moderate COPD patients. Calibration bar represents 20 μm. Graphical representation of the results are shown in the right hand panels. (JPG 332 kb

Additional file 4: Figure S2. of Reduced suppressive effect of β2-adrenoceptor agonist on fibrocyte function in severe asthma

Effect of salmeterol on NANT cell apoptosis. NANT cells from healthy subjects (n = 6) were treated with salmeterol (10−8 M) for 3 days and the percentage of live, and early and late apoptotic NANT cells was determined. Bars represent mean ± SEM. (PDF 173 kb

Additional file 1: of Reduced suppressive effect of β2-adrenoceptor agonist on fibrocyte function in severe asthma

Supplementary Materials and Methods. (DOCX 19 kb

Additional file 3: Figure S1. of Reduced suppressive effect of β2-adrenoceptor agonist on fibrocyte function in severe asthma

α–smooth muscle actin-positive cells also express both collagen I and CD45. NANT cells from one healthy subject were harvested after 3 days in culture and stained with antibodies for collagen I (Col I), α- smooth muscle actin (SMA) and CD45 or their respective IgG isotype controls. (A and C). The percentage of Col I+/α-SMA+ (B) and CD45+/ α-SMA+ cells (D) was determined by flow cytometry. (PDF 112 kb

Additional file 2: Figure S3. of Reduced suppressive effect of β2-adrenoceptor agonist on fibrocyte function in severe asthma

Number of fibrocytes and differentiating fibrocytes in NANT cells after culture in the absence or presence of salmeterol. The number of fibrocytes (% Col I+/CD45+; A) and differentiating fibrocytes (% α-SMA+; B) within the NANT cell population, were determined in the NANT cells from healthy subjects (n = 8–9) and patients with non-severe (n = 7) or severe asthma (n = 7–9) at day 0 (0 d) or after 3 days (3 d) in the absence or presence of salmeterol (10−9- 10−7 M). Data points represent mean ± SEM. * p < 0.05 and ** p < 0.01 versus vehicle-treated cells for each group. ### p < 0.001 versus healthy group and $$$ p < 0.001 versus non-severe asthma group. (PDF 478 kb

Additional file 5: Figure S4. of Reduced suppressive effect of β2-adrenoceptor agonist on fibrocyte function in severe asthma

Effect of ICI-118,551 on salmeterol-mediated reduction in fibrocyte number and differentiation. NANT cells from healthy subjects (n = 6) were treated with salmeterol (10−8 M) in the presence or absence of ICI-118,551 (10−5 M) for 3 days. The number of fibrocytes (Col I+/CD45+ cells; A) and differentiating fibrocytes (α-SMA+ cells; B) was determined. Bars represent mean ± SEM.* p < 0.05 and ** p < 0.01. (PDF 69 kb

MOESM2 of Proteomic analysis of sputum reveals novel biomarkers for various presentations of asthma

Additional file 2: Figure S1. High expression of secreted protein in CA, CVA, or CTVA patients. AGT (A), ANXA1 (B), APOA1 (C), B2M (D), C5 (E), CHI3L1 (F), CTSB (G), FGA (H), FGB (I), FN1 (J), HPX (K), IL1RN (L), ITIH4 (M), ORM1 (N), PRG2 (O), and SERPINF (P). Mean values are represented as horizontal bars. *P < 0.05, **P < 0.01, ***P < 0.001; CA, classic asthma; CVA, cough-variant asthma; CTVA, chest tightness variant asthma

Additional file 3: of A computational framework for complex disease stratification from multiple large-scale datasets

Table S7. Estimated accuracy and standard deviation of the RFE procedure. Table S8. Accuracy and Kappa values of the Random Forest models in the training set. Table S9. Performances values for the Random Forest model in the testing set. Figure S11. Relative importance of the top 20 predictors building the final model of the RF. The importance axis is scaled, with the mRNA expression of CD3D scaled to 100% and the methylation state of POLA2 to 0% (not shown). (DOCX 18 kb

Additional file 4: of A computational framework for complex disease stratification from multiple large-scale datasets

DIABLO sPLSDA model results. (DOCX 18966 kb