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3 protocols using damascenone

1

Chemical Compound Sourcing for Research

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Fructose (99%) and tannic acid (95%) were purchased from J.T. Baker (Avantor; Radnor, PA, USA). Tartaric acid (99.7%) was provided by Chem-Lab (Eernegem, West-Vlaanderen, Belgium), and caffeine (99.2%) by ACEF (Piacenza, Italy). 2-Phenylethanol (≥99%), citral (95%), linalool (97%), 1-octen-3-one (96%), cis-3-hexen-1-ol (≥98%), ethyl butyrate (≥99%), damascenone (1.1–1.4 wt.%), benzaldehyde (≥99.5%), isoamyl acetate (≥95%), gamma-dodecalactone (≥97%), sotolone (≥97%), 4-ethylguaiacol (≥98%), 4-ethylphenol (99%), eucalyptol (99%), furaneol (≥98%), ethyl caproate (≥99%), eugenol (≥98%), citronellol (95%), phenylacetaldehyde (≥95%), furfuryl acetate (≥98%), 2,4,6-trichloroanisole (99%), 2-methyl-1-propanol (99.5%), methanethiol (≥98%) were all provided by Sigma-Aldrich (St. Louis, MO, USA). Ethanol (food grade, 70%) was supplied by ITW Reagents (Milano, Italy). Tanin VR colour, Tanin VR grape, Tanin plus, Tanin galalcool were all purchased from Laffort (Bordeaux, France).
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2

Primary Human Airway Cell Culture Protocol

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Cellgro DMEM:F12 was from Mediatech (Manassas, VA). Lechner and LaVeck basal media (LHC), Hanks’ Balanced Saline Solution, glutamax, penicillin, and streptomycin were from Life Technologies (Carlsbad, CA). Fibronectin, type I collagen, and Nu-Serum™ were from Becton-Dickinson (Franklin Lakes, NJ). Minimum Essential Medium with Earle’s salts (MEM), Fetal Bovine Serum (FBS), 2,5-dimethylpyrazine, amiloride, damascenone, forskolin, linalool, α-ionone, ethyl maltol, furaneol and vanillin were from Sigma-Aldrich (St. Louis, MO). CFTR-172 inh and 8-bromo-cAMP were from Tocris Bioscience (Bristol, UK). Semipermeable filters were Corning Costar 6.5 mm Transwell® with 0.4 μm Pore Polyester Membrane Insert, sterile (Lowell, MA). All other chemicals were from Sigma-Aldrich or Fisher Scientific (Pittsburgh, PA).
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3

Purified Compounds for ATP Synthase Assay

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Safranal with ≥88% purity (W338907-25G-K, Sigma-Aldrich), thymol with 99% purity (150335000, Acros-Organics), carvacrol with >95% purity (C0026, Tokyo Chemical Industry), damascenone with Analytical Standard purity (41163-250MG, Sigma-Aldrich), cuminol with 97% purity (187801000, Acros-Organics), TMCHD with ≥98% purity (L10199, Alfa Aesar), IPBB with 97% purity (187801000, Acros-Organics), and TBP with 97% purity (108000050, Acros-Organics) were used in this study. Many were a liquid at room temperature and were dissolved in dimethyl sulfoxide (DMSO) to obtain the desired working solution. In ATPase assays, the maximal volume of DMSO used was 3.64%. In this study and previous studies, we noted that up to 40% DMSO by itself has no effect on membrane-bound F1Fo of E. coli ATP synthase [61 (link), 62 (link)]. All other chemicals used in this study were ultrapure, analytical grade purchased from Sigma-Aldrich Chemical Company or Fisher Scientific Company.
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