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. 2021 Mar 23;11:6655. doi: 10.1038/s41598-021-85742-0

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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MMP activity and IL-6 level in the medium of JIMT-1 and HDF 3D mono-cultures and of JIMT1/HDF 3D co-cultures. The cultures were incubated in the absence (control) or presence of 5 ng/ml TGF-β1. After 14 days of incubation, the medium was collected for analysis of MMP activity (A–C) and the concentration of IL-6 (D–E). (A–C) The MMP activity was determined in 50 μl of medium by an assay based on the formation of a fluorescent product produced by MMP enzyme activity. (D–F) The level of IL-6 in the medium was determined by an ELISA assay. (G–L) Following the collection of the culture media, the cells were fixed with 3.7% formaldehyde and stained to visualize IL-6 (green), vimentin (red), and cell nuclei (blue). The images show a single confocal microscopy plane taken in the centre of the cultures. Each column represents mean ± SEM (n = 5). Students t-test, *P < 0.05. Scale bar is 50 μm.