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. 2020 Jul 21;10:12120. doi: 10.1038/s41598-020-68995-z

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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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IL-10 modulates TLR2 innate signaling and OxPCs-mediated changes in global ischemia–reperfusion (I/R) injury. Hearts were subjected to global I/R with and without IL-10 as described in the methods. For IL-10 treatment, 10 ng/ml of IL-10 was added for 40 min during the reperfusion phase. (A) Total, fragmented and non-fragmented OxPCs were measured using mass-spectrometer for each fraction and their levels in I/R hearts were normalized compared with those in the presence of IL-10. (B) Non-oxidized lipids (PC Phosphatidylcholine and SM Sphingomyelin). (C) Western blotting was used to determine protein expression of mLOX1 and LOX1 post-I/R. (D) Expression of TLR2 was detected in the myocardium using immunofluorescence. Percentage of mean fluorescence intensity (% MFI) was calculated from three different heart sections and counted in 10 different fields after TLR2 staining. Upper panel is showing for Hematoxylin and Eosin (H/E) staining. (E) Western blots were probed with antibody for SREBP 1c^ser372 . GAPDH was used as a loading control. Histogram (E, lower panel) is representative of 3 independent hearts probed for SREBP1c^ser372. (F) TGF-β receptor II (TGF-βRII) expression was also studied in tissue sections using ABC peroxidase staining. Sections were probed with specific TGF-βRII antibody followed by diaminobezidine (DAB) staining.