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. 2021 Mar 18;11:6268. doi: 10.1038/s41598-021-85241-2

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© The Author(s) 2021

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 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/.

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Analysis of adhesion molecules expressed in vascular endothelial cells stimulated by LPS and MP. (a) Protein expression levels of E-selectin, VCAM-1, and ICAM-1 in HUVEC at 6 h after stimulation analyzed by western blotting. Blot was cut horizontally and immunoblotting was performed for each section. (b) Quantification of E-selectin, VCAM-1, and ICAM-1 band intensity normalized to β-actin in HUVEC. Data are means ± standard error (n = 3, each group); *P < 0.05. (c) Induction of SELE, VCAM1, and ICAM1 gene expression in HUVEC 6 h after stimulated by 1.0 µg/mL of LPS and 100 µg/mL of MP was analyzed by quantitative real-time PCR. Data are normalized relative to ACTB mRNA levels. Data are means ± standard error (n = 3, each group); *P < 0.05. (d) Protein expression levels of E-selectin, VCAM-1, and ICAM-1 in HHSEC at 6 h after stimulation analyzed by western blotting. Blot was cut horizontally and immunoblotting was performed for each section. (e) Quantification of E-selectin, VCAM-1, and ICAM-1 band intensity normalized to β-actin in HHSEC. Data are means ± standard error (n = 3, each group); *P < 0.05. (f) The phosphorylation of Nf-κB, JAK1, and STAT3 was analyzed by western blotting with antibodies targeting pNf-κB/Nf-κB, pJAK1/JAK1, and pSTAT3/STAT3 at 30 min after the stimulation. Blot was cut horizontally and immunoblotting was performed for each section. (g) ELISA analysis of IL-6 concentration levels in conditioned medium 6 h after stimulated by 1.0 µg/mL of LPS and 100 µg/mL of MP. Data are means ± standard error (n = 3, each group); *P < 0.05.