Fig. 1.

Anandamide (AEA) is released by vascular cells and attenuates induction of a subset of IL-1β responsive genes in HAoSMC. a Cellular concentration of the indicated endocannabinoids as determined by LC–MS/MS from murine aortic rings treated with TNFα and IL-1β 10 ng/mL each, 4 h or with control (PBS with 0.1% BSA). N = 4–6, unpaired t test. b Heatmap showing Z score of restricted to IL-1β-induced genes with expression significantly altered by 100 nmol/L, 150 min AEA and subsequently 10 ng/mL IL-1β for 90 min in HAoSMC. N = 3 RT-qPCR of CCL2 performed from human aortic (c) and mouse aortic (d) smooth muscle cells and murine aortic rings (e) in response to the stimuli indicated. Cells treated with 100 nmol/L AEA for 150 min (for tissue: overnight) and subsequently stimulation with 10 ng/mL cytokine for 90 min. N = 5–10 f Protein levels of CCL2 as determined by Western blot analysis after treatment with 100 nmol/L AEA for at least 1 h and subsequently 10 ng/mL IL-1β for 3 h (normalized to β-tubulin, N = 3) or ELISA N = 5. Ordinary one-way ANOVA with Tukey’s multiple comparison post-hoc test. If not normally distributed nonparametric ANOVA with Kruskal–Wallis test and Dunn’s multiple comparison post-hoc test