Endothelial CCL5 expression. (A) Detection of C-C chemokine ligand 5 (CCL5) mRNA in HMVECs purified from cocultures as indicated. Mean ± SEM; n = 3. As positive controls, HMVECs were stimulated with TNFα (10 ng/mL) and interferon γ (1 ng/mL). (B) CCL5 protein levels secreted by cocultured HMVECs and measured by cytometric bead array. The de novo CCL5 production was measured in the newly added medium after 12 hours. Mean ± SEM; n = 6-8. (C) Adherence of monocytes to HMVECs activated by different coculture conditions. Monocytes were allowed to adhere to HMVEC layers for 2 hours. Mean ± SEM; n = 4. (D) Migration of monocytes toward cocultured HMVECs. Migration of monocytes was determined in the presence of a CCL5-blocking antibody or of a corresponding isotype control IgG. Mean ± SEM; 3 independent experiments. Statistical significance was determined ANOVA with Bonferonni multiple comparison test; *P < .05, **P < .01, ***P < .001.