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. 2019 Apr 1;9:5399. doi: 10.1038/s41598-019-41782-1

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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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Sequential co-incubation of catK then catL with collagen gels sustained active catK, while bi-directional cannibalism of catK and catL occurred during concurrent co-incubation. Zymography and Western blots for catK and catL were conducted to measure the amount of active catK and catL remaining after incubation with collagen gels. CatK and catL were active (A) after 8 hr incubation with collagen as indicated by zymography in the soluble and pellet fractions. Significantly more catK was active when collagen was sequentially co-incubated with catK then catL (→) than concurrent co-incubation (+). (B) More catL was active when collagen was primed with catK (→) than concurrent co-incubation with catK (+). (C) CatK Western blot detected no catK in the soluble fraction, but in the pellet fraction less total catK remained after concurrent co-incubation with catL (+), suggesting catL was hydrolyzing catK (D,E). By Western, less total catL was present when co-incubated with catK, even when sequentially co-incubated with catK (→), suggesting catK may also have been hydrolyzing catL. (D,F) n = 3, *indicates statistical significance, p < 0.05 as determined by a one-way ANOVA. Error bars represent SEM.