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. 2021 Aug 20;23:218. doi: 10.1186/s13075-021-02588-7

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© The Author(s) 2021

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 6 — TSG6-mediated intermolecular crosslinking of HA. Size exclusion chromatography-multi-angle light scattering (SEC-MALS) was used to determine how the quaternary structure of monodisperse, 150 kDa HA was altered in the presence of TSG-6 and inter-alpha-inhibitor protein (IαIp), with one replicate (n = 1) for each treatment condition. A Control solution: HA and IαIp. Two significant peaks (HA and IαIp) were observed at their calculated MW of 141 kDa and 154 kDa, respectively. B, C HA, TSG-6, and IαIp. C was created by re-scaling the MW y-axis of B to show the high MW of HA aggregates. The light scattering (LS) shows 2 peaks with higher MW of ~ 220 kDa (equivalent to the MW of HC-HA complex, HC + HA = 75 kDa + 150 kDa) a weighted average of ~ 3288 kDa. The bold, dotted blue line indicates the calculated molecular weight across each peak, corresponding to MW (kDa) on the right y-axis. The dashed blue line represents the expected MW of 150 kDa for the monodisperse HA solution. D Viscosity increased in the TSG6-mediated HC-HA crosslinking reaction mixture (HA + IαI + TSG6) as compared to all other 3 controls. Data are displayed as scatter plots representing the mean ± SD (n = 5 technical replicates)