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. 2021 Jan 21;11:1949. doi: 10.1038/s41598-020-80102-w

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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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Adhesion molecule expression by cytokine-activated endothelial cells. (a) After 4 h activation with TNFα (20 ng/mL) or IL-1β (20 ng/mL), stimulated endothelial cells were lysed in RLT buffer, and mRNA for immune response genes was measured by nanostring. Absolute mRNA counts for endothelial adhesion molecule genes are presented in the heat map. (b, c) After 4 h or 18-24 h, stimulated endothelial cells were detached and TNFα (b) and IL-1β (c) induced cell surface E-selectin was measured by flow cytometry. Results are presented as fold increase in the median fluorescence intensity of cell surface E-selectin, normalized within each experiment to the untreated condition for each cell type.(d, e) After 4 h or 18-24 h, stimulated endothelial cells were detached and TNFα (d) and IL-1β (e) induced cell surface ICAM-1 was measured by flow cytometry. Results are presented as fold increase in the median fluorescence intensity of cell surface ICAM-1, normalized within each experiment to the untreated condition for each cell type. (f, g) After 4 h or 18-24 h, stimulated endothelial cells were detached and TNFα (f) and IL-1β (g) induced cell surface VCAM-1 was measured by flow cytometry. Results are presented as fold increase in the median fluorescence intensity of cell surface VCAM-1, normalized within each experiment to the untreated condition for each cell type. (TNFα and IL-1β 4 h, n = 4; 18-24 h, n = 5–6 donors per endothelial cell type). #p < 0.1, *p < 0.05, **p < 0.001 comparing HAEC and HCAEC to HLSEC by two way ANOVA followed by uncorrected Fisher’s LSD. (h) mRNA and (i) fold increase in the MFI of cell surface expression of VCAM-1 was measured on aortic HAEC and liver HLSEC endothelial cells stimulated with IL-4 (20 ng/mL) for 4 h or 24 h (n = 3 donors per endothelial cell type). Results are presented as mean ± SEM. #p < 0.1 comparing HAEC to HLSEC, by two way ANOVA and uncorrected Fishers LSD.

Adhesion molecule expression by cytokine-activated endothelial cells. (a) After 4 h activation with TNFα (20 ng/mL) or IL-1β (20 ng/mL), stimulated endothelial cells were lysed in RLT buffer, and mRNA for immune response genes was measured by nanostring. Absolute mRNA counts for endothelial adhesion molecule genes are presented in the heat map. (b, c) After 4 h or 18-24 h, stimulated endothelial cells were detached and TNFα (b) and IL-1β (c) induced cell surface E-selectin was measured by flow cytometry. Results are presented as fold increase in the median fluorescence intensity of cell surface E-selectin, normalized within each experiment to the untreated condition for each cell type.(d, e) After 4 h or 18-24 h, stimulated endothelial cells were detached and TNFα (d) and IL-1β (e) induced cell surface ICAM-1 was measured by flow cytometry. Results are presented as fold increase in the median fluorescence intensity of cell surface ICAM-1, normalized within each experiment to the untreated condition for each cell type. (f, g) After 4 h or 18-24 h, stimulated endothelial cells were detached and TNFα (f) and IL-1β (g) induced cell surface VCAM-1 was measured by flow cytometry. Results are presented as fold increase in the median fluorescence intensity of cell surface VCAM-1, normalized within each experiment to the untreated condition for each cell type. (TNFα and IL-1β 4 h, n = 4; 18-24 h, n = 5–6 donors per endothelial cell type). #p < 0.1, *p < 0.05, **p < 0.001 comparing HAEC and HCAEC to HLSEC by two way ANOVA followed by uncorrected Fisher’s LSD. (h) mRNA and (i) fold increase in the MFI of cell surface expression of VCAM-1 was measured on aortic HAEC and liver HLSEC endothelial cells stimulated with IL-4 (20 ng/mL) for 4 h or 24 h (n = 3 donors per endothelial cell type). Results are presented as mean ± SEM. #p < 0.1 comparing HAEC to HLSEC, by two way ANOVA and uncorrected Fishers LSD.