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. 2020 Feb 13;10:2544. doi: 10.1038/s41598-020-59290-y

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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E-liquids increased mucin production in HMEECs. (A) HMEECs were exposed to e-liquids at the IC₅₀ concentration for 24 h (PG/VG: 4.5%, Tobacco: 3.3%, and Menthol: 1.5%), and quantitative RT-PCR was performed to investigate the expression levels of mucin genes such as MUC4, MUC5AC, and MUC5B. Tobacco- and menthol-flavored e-liquids significantly increased the mRNA expressions of MUC4 and MUC5AC. Data were obtained from three independent experiments and the error bars indicate the mean ± SD. *p < 0.05, **p < 0.01, and ***p < 0.001 compared to in the corresponding control. (B) Cells were treated with e-liquids at the IC₅₀ concentration for 24 h (PG/VG: 4.5%, Tobacco: 3.3%, and Menthol: 1.5%). Confocal microscopic images showing MUC5AC expression (green) in HMEECs as detected by immunofluorescence staining. MUC5AC expression in cells was higher in e-liquid-treated groups than in the control group. Nuclei were counterstained with DAPI (blue).