Fig. 6
High-CX3CL1 expression regulates the luminal retention of proangiogenic monocytes. a Scheme of proangiogenic monocyte transmigration map indicating how cytokines and combinations of factors affect proangiogenic monocyte transmigration. b Screening by qPCR for differential mRNA expression by endothelial cells of molecules involved in leucocyte recruitment including capture, crawling and transmigration, after indicated stimulations. Normalisation was performed with housekeeping gene mRNA levels namely ACTB and GAPDH. The level in inflammation is expressed as the fold-change of expression level compared to non-stimulated endothelial cells. The horizontal dashed line (red) indicates baseline level. n = 3 technical replicates, representative of three independents experiments, the data are mean ± s.e.m. c Protein expression of CX3CL1 by endothelial cells upon activation with different cytokines and combination of factors. d Western blot showing efficient downregulation of CX3CL1 expression in HUVEC after transfection with a plasmid expressing CX3CL1-targeting shRNA. e Effect of CX3CL1 downregulation on transmigration of monocyte subsets through TNF or TNF + IFNγ-activated endothelium. n = 4 independent experiments with monocytes from four different donors, the data are presented as mean ± s.e.m., *p < 0.05; ****p < 0.0001 in ANOVA test
