Reduced expression of chemokines in anti-IL1RAP-treated mice. RNA was isolated from BCAs from HCD-fed Apoe^−/− mice treated with anti-IL1RAP antibodies or isotype control (Ctrl IgG) (pooled samples, n = 2/pool; n = 7/group). Real-time PCR quantification of relative expression (2^−ΔΔCt) of (A) adhesion molecules, (B) chemokines, and (C) inflammatory cytokines. (D) Dot plot of gene expression of chemokines and adhesion molecules in human carotid artery plaques. (E) BMDMs from wild-type (C57Bl/6) mice and a mouse fibroblast cell line (NIH3T3) were stimulated with either IL-1α, IL-1β, IL-33, or IL-36 in the presence of anti-IL1RAP antibody or isotype control (Ctrl IgG). (F and G) Expression of IL1RAP on BMDMs and NIH3T3 fibroblasts determined by flow cytometry. (H) Quantification of release of CXCL1 by NIH3T3 fibroblasts under each treatment condition (n = 6). Quantification of release of CXCL1 by (I) BMDMs (n = 3) stimulated with IL-1α, IL-1β, or IL-33 or (J) BMDMs (pre-treated with GM-CSF/TGF-β; n = 5) stimulated with IL-36 (IL-36α, IL36-β, and IL-36γ). (A–C) Bars denote median, analysed with Mann–Whitney U test (*P = 0.0262). (H–J) Bars denote mean.