Impact of myeloid Cnr1 deficiency on circulating leucocyte counts, chemokine receptor expression, and arterial recruitment. (A, B) Number of circulating monocytes in male (7–19) and female (5–23) Apoe^−/−LysM^Cre and Apoe^−/−LysM^CreCnr1^flox/flox mice assessed by flow cytometry. (C) Flow cytometric analysis of colony-stimulating factor 1 receptor (CSF1R) expression on circulating monocytes after 4 weeks of WD (n = 6–18). (D, E) Flow cytometric analysis of CCR1 and CCR5 surface expression on circulating monocytes after 4 weeks of WD (n = 6–8). (F) Microscopy images and quantification of CFSE-labelled monocytes (green) recruited into aortic root lesions 48 h after injection into male Apoe^−/− mice on WD for 4 weeks (n = 3–4), Nuclei were stained with Hoechst 33342 (blue); scale bar: 100 μm (left) and 10 µm (right). (G) Flow cytometric analysis of male BMDM treated with vehicle or 10 nM oestradiol (E2) for 24 h (n = 6). (H, I) Gene expression levels in untreated male and female BMDMs (n = 5–12). (J, K) Flow cytometric analysis of male BMDM from Apoe^−/− mice treated with vehicle, 10 µM AM281 or 1 µM ACEA for 24 h (n = 4–6). (L) Gene expression levels in male Apoe^−/− BMDMs treated with vehicle or 10 µM AM281 for 8 h (n = 5–6). Each dot represents one biologically independent mouse sample and all data are expressed as mean ± S.E.M. Two-sided unpaired Student’s t-test (A–C, F, H, I, and L), one-way ANOVA followed by Tukey test (J and K), two-way ANOVA followed by Tukey test (D, E, G). Male and female were analysed independently (A–I).