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. 2019 Sep 25;9:13905. doi: 10.1038/s41598-019-50307-9

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© The Author(s) 2019

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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MMC reduces αSMA and fibronectin localization and expression in HCFs and conditioned media from MMC treated HCLE cells blocks TGFβ1-induced collagen deposition. (A) As indicated in the schematic, control and MMC-treated HCFs were grown in HCF media and in media supplemented with vitamin C and TGFβ1 for 48 hours. Cells were fixed, permeabilized and stained with antibodies against αSMA (red) and collagen I (green) and TN-C (red) and fibronectin (green). In addition, nuclei were stained with DAPI (blue). Data show that 48 hours after MMC treatment, αSMA expression and FN fibril formation in HCFs is reduced. The magnification bar = 40 μm. (B) To determine whether exposure to proteins secreted by MMC-treated HCLE cells impacts collagen deposition by HCFs, E-CMC and E-CMM was added to HCF cultures and collagen deposition assessed. Data show that the addition of vitamin C and TGFβ1 to E-CMM did not induce collagen deposition but addition of vitamin C and TGFβ1 to E-CMC increased collagen deposition by HCFs significantly.