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. Author manuscript; available in PMC: 2022 Jun 23.
Published in final edited form as: Methods Mol Biol. 2022;2419:169–182. doi: 10.1007/978-1-0716-1924-7_10

Table 3.

Methods for assessing endothelial cell activation in vitro

Technique Description PMID
Monocyte adhesion assay Used to measure adhesiveness of blood monocytes into endothelial cell layer. 26733204, 26085094, 29371247, 27127201
Trans-well migration assay Commonly used to study the migratory response of endothelial cells toward a chemoattractant. 29511499
Real-time polymerase chain reaction (RT-PCR) Used to examine mRNA expression levels of adhesion molecules such as ICAM-1, VCAM-1, and E-selectin. 29371247
Western blotting Used to examine protein levels of adhesion molecules such as ICAM-1, VCAM-1, and E-selectin. 29371247, 25705917
Flow cytometry analysis Used to examine expression of adhesion molecules such as ICAM-1 and VCAM-1 as well as monocyte migration. 25705917, 26085094, 26733204
Cytokine array Used to determine cytokine and chemokine expression in activated endothelial cells. 26085094, 25705917
RNA sequencing Provides information on the expression levels of adhesion molecules and other endothelial cell activation genes. 29769317, 29371247
Microarray analysis Used to examine the expression levels of adhesion molecules and cytokines/chemokines. 26733204, 27127201, 29371247
Endothelial cell biology PCR array Used to examine expression of 84 endothelial cell-related genes. 27127201
Enzyme-linked immunosorbent assay (ELISA) Used to determine protein levels of different cytokine/chemokines released from activated endothelial cells. 26085094
FAM-FLICA caspase-1 assay Utilizes the fluorescent inhibitor probe FAM-YVAD-FMK to label active caspase-1 enzyme in living cells (the inflammatory sites of the endothelium). 25739025

ICAM-1 intercellular adhesion molecule-1, VCAM vascular cell adhesion molecule-1