Fig. 2. Macrophage-derived 27HC drives vascular inflammation and promotes atherosclerosis via endothelial ERα.

a Representative still images of leukocyte-endothelial cell adhesion evaluated by intravital microscopy in ERα^fl/fl and ERα^∆EC mice injected with vehicle versus 27HC (20 mg/kg BW) daily for 3 days. b Summary data for leukocyte velocity in (a), n = 11-12. c–f ApoE^-/- background ERα^fl/fl and ERα^ΔEC mice were placed on an atherogenic diet for 8 weeks and administered vehicle versus 27HC, and samples were obtained. Representative lipid-stained en face images of aortas (c), and lesion areas (d, percent of total surface area). N = 13 and 10 for ERα^fl/fl given vehicle versus 27HC, respectively, and n = 10 and 12 for ERα^ΔEC given vehicle versus 27HC, respectively. Representative lipid and haematoxylin-stained aortic root sections (e), and lesion areas (f, n = 10). g–o, Atherosclerosis and related parameters in bone marrow transplant experiments performed employing apoE^-/- background cyp27a1^fl/fl versus cyp27a1^ΔMAC donors and apoE^-/- background ERα^fl/fl versus ERα^ΔEC recipients. g Schematic of donors and recipients. Representative lipid-stained en face images of aortas (h) and lesion areas (i, n = 11-12). Representative lipid and haematoxylin-stained aortic root sections (j), and lesion areas (k, n = 11-12). Representative immunohistochemistry images of Mac2 staining in aortic root (l; nuclei DAPI stained) and quantification of Mac2 positive staining (m; expressed per unit lesion area, n = 8−10). Immunohistochemical determinations of endothelial ICAM-1 and VCAM-1 abundance with representative images (n) and quantification (o, n = 8-9). Arrows indicate endothelium. Scale bar equals 100 um (e, j) or 50 um (l, n). Data are mean ± SEM. In (b) and (d), P values shown are by Krusakal-Wallis with Dunn’s post hoc testing. In all other graphs (f, i, k, m, o), P values shown are by ANOVA with Tukey’s post-hoc testing. Source data are provided as a Source Data file.