A–h) Comparison of the effects of cigarette smoke extract (CSE), electronic cigarette (eCig) vapour and IQOS aerosol exposure on a–d) human airway epithelial.

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a–h) Comparison of the effects of cigarette smoke extract (CSE), electronic cigarette (eCig) vapour and IQOS aerosol exposure on a–d) human airway epithelial (Beas-2B) and e–h) human airway smooth muscle (ASM) cells. a and e) show the release of CXCL8 from ... a–h) Comparison of the effects of cigarette smoke extract (CSE), electronic cigarette (eCig) vapour and IQOS aerosol exposure on a–d) human airway epithelial (Beas-2B) and e–h) human airway smooth muscle (ASM) cells. a and e) show the release of CXCL8 from Beas-2B and ASM cells. The concentration of CXCL8 in supernatant from Beas-2B and ASM cells after 72 h of stimulation with CSE, eCig vapour or IQOS aerosol exposure was measured using ELISA. Deposition of b and f) collagen I alpha 1 (COL1A1) and c and g) fibronectin from Beas-2B and human ASM cells after 72 h of stimulation with CSE, eCig vapour or IQOS aerosol exposure was measured using extracellular matrix (ECM) ELISA at an absorbance of 450 nm and 570 nm, respectively. d and h) The level of glycolysis was determined in Beas-2B and ASM cells using a seahorse analyser, and extracellular acidification rate (ECAR); an index of glycolysis was measured after 72 h of stimulation with CSE, eCig vapour or IQOS aerosol exposure. Data are presented as mean±sem (n=5–7). i–n) The effect of CSE, eCig and IQOS exposure on cellular toxicity and respiration. The cell viability (i and l), the lactate dehydrogenase (LDH) release (j and m) and the mitochondrial respiration (k and n) from Beas-2B and human ASM cells was measured using Thaizolyl blue tetrazolium bromide (MTT) and LDH assays at an absorbance of 570 nm and 490 nm, respectively. k and n) Mitochondrial respiration was measured in Beas-2B and ASM cells using a mito-stress kit on a seahorse analyser (Agilent Technologies Inc., Santa Clara, CA, USA), and proton leak was measured as oxygen consumption rate shown as fold change to control. Cells were stimulated with serial dilution of CSE, eCig vapour or IQOS aerosol for 72 h (n=5). Data are presented as mean±sem. A one-way ANOVA plus Bonferroni post-test was used to determine statistical significance. *: p<0.05 compared with control; **: p<0.01 compared with control; ***: p<0.001 compared with control. Sukhwinder Singh Sohal et al. ERJ Open Res 2019;5:00159-2018 ©2019 by European Respiratory Society