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. 2020 May 19;10:8250. doi: 10.1038/s41598-020-64931-3

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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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Collagen solubilization assay. (Panel A) Experimental outline of the protocol; collagen type 1 was polymerized in the presence of neutralizing buffer, followed by treatment with the test reagents, and incubated overnight at room temperature; the supernatant from each tube was run on SDS-PAGE to assess the released monomeric soluble collagen. A representative image showing decreased opacity of the polymerized collagen in the presence of CBD as compared to the control tube (minus CBD); (Panel B) Representative SDS-PAGE gel of the assay; (Panel C) Upon treatment with CBD/RCBD, polymerized collagen releases the monomeric form of (soluble) collagen, seen to be increased in samples treated with reagents (indicated by increased band intensity of α1 and α2 chains of collagen), as compared to untreated sample of collagen (control). Negative control: 20 μM CK (cysteine knot domain in the C-terminal region of CTGF); Data represents the mean (± standard deviation, SD) of five independent experiments (*p < 0.05, significant; NS, Not Significant).