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. 2020 Nov 9;22:265. doi: 10.1186/s13075-020-02360-3

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© The Author(s) 2020

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. 4 — Intra-cellular signaling pathways in HUVECs stimulated with SSc-ICs, PAPS-ICs, SLE-ICs, or NHS-ICs. Endothelial cells were incubated with SSc-ICs, PAPS-ICs, SLE-ICs, or NHS-ICs (1:2 dilution). IL-1β (50 U/ml) was used as a positive control. Results are expressed as the ratio of phosphorylated to non-phosphorylated forms, evaluated using the ImageJ software. Western Blotting images are representative of a single experiment. pNFκB: phosphorylated NFκB; p38MAPK: phosphorylated p38MAPK; pp46SAPK-JNK: phosphorylated p46SAPK-JNK; pp54SAPK-JNK: phosphorylated p54SAPK-JNK; pAkt: phosphorylated Akt. Histograms represent mean + standard error of the mean (SEM). *p < 0.01; **p < 0.001; ***p < 0.0001 versus medium. a pNFκB/NFkB. IL-1β [7.75 ± 0.49] versus medium [1.02 ± 0.05] (p < 0.0001) versus NHS-ICs (p < 0.0001). ATA-ICs [5.00 ± 0.55] versus medium (p < 0.0001) and versus NHS-ICs [22.75 ± 7.59] (p < 0.0001). ACA-ICs [3.00 ± 1.39] versus medium (p < 0.01) versus NHS-ICs (p = N.S.). ARA-ICs [2.17 ± 1.55] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). Anti-Th/To-ICs [2.98 ± 1.75] versus medium (p < 0.01) versus NHS-ICs (p = N.S.). PAPS-ICs [2.13 ± 0.26] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). SLE-ICs [1.65 ± 0.13] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). NHS-ICs [1.25 ± 0.21] versus medium (p = N.S.). b pp38/p38. IL-1β [31.98 ± 9.37] versus medium [1.4 ± 0.4] (p < 0.0001) versus NHS-ICs (p < 0.0001). ATA-ICs [26.75 ± 7.3] versus medium (p < 0.0001) and versus NHS-ICs [22.75 ± 7.59] (p < 0.001). ACA-ICs [22.43 ± 9.57] versus medium (p < 0.001) versus NHS-ICs (p < 0.01). ARA-ICs [26.15 ± 5.27] versus medium (p < 0.001) versus NHS-ICs (p < 0.001). Anti-Th/To-ICs [24.65 ± 12.77] versus medium (p < 0.001) versus NHS-ICs (p < 0.001). PAPS-ICs [21.93 ± 7.77] versus medium (p < 0.001) versus NHS-ICs (p < 0.01). SLE-ICs [11.63 ± 9.14] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). NHS-ICs [3.2 ± 0.85] versus medium (p = N.S.). c pp46SAPK-JNK/p46SAPK-JNK. IL-1β [1.98 ± 0.17] versus medium [1.03 ± 0.05] (p < 0.0001) versus NHS-ICs (p < 0.0001). ATA-ICs [1.85 ± 0.19] versus medium (p < 0.0001) and versus NHS-ICs [22.75 ± 7.59] (p < 0.0001). ACA-ICs [1.65 ± 0.13] versus medium (p < 0.001) versus NHS-ICs (p < 0.0001). ARA-ICs [1.60 ± 0.48] versus medium (p < 0.01) versus NHS-ICs (p < 0.001). Anti-Th/To-ICs [1.23 ± 0.22] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). PAPS-ICs [1.22 ± 0.09] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). SLE-ICs [1.13 ± 0.19] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). NHS-ICs [0.88 ± 0.32] versus medium (p = N.S.). d pp54SAPK-JNK/p54SAPK-JNK. IL-1β [20.88 ± 1.56] versus medium [1.05 ± 0.06] (p < 0.0001) versus NHS-ICs (p < 0.0001). ATA-ICs [7.30 ± 0.63] versus medium [21.75 ± 6.24] (p < 0.0001) and versus NHS-ICs [22.75 ± 7.59] (p < 0.0001). ACA-ICs [3.75 ± 0.59] versus medium (p < 0.001) versus NHS-ICs (p < 0.01). ARA-ICs [2.52 ± 0.41] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). Anti-Th/To-ICs [2.02 ± 0.60] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). PAPS-ICs [3.98 ± 0.24] versus medium (p < 0.001) versus NHS-ICs (p < 0.01). SLE-ICs [3.05 ± 2.17] versus medium (p < 0.01) versus NHS-ICs (p = N.S.). NHS-ICs [1.73 ± 0.28] versus medium (p = N.S.). e pAkt/Akt. IL-1β [1.90 ± 0.39] versus medium [1.02 ± 0.05] (p < 0.01) versus NHS-ICs (p < 0.01). ATA-ICs [1.50 ± 0.24] versus medium [21.75 ± 6.24] (p = N.S.) and versus NHS-ICs [22.75 ± 7.59] (p = N.S.). ACA-ICs [1.80 ± 0.48] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). ARA-ICs [1.70 ± 0.14] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). Anti-Th/To-ICs [1.90 ± 0.42] versus medium (p < 0.01) versus NHS-ICs (p < 0.01). PAPS-ICs [2.02 ± 0.59] versus medium (p < 0.001) versus NHS-ICs (p < 0.01). SLE-ICs [1.42 ± 0.65] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). NHS-ICs [1.08 ± 0.10] versus medium (p = N.S)