Fig. 3. HSC-derived arterial macrophages.

a, b Flow-cytometry analysis of arterial macrophages of Flt3^CreRosa26^eYFP mice (gated on single CD45⁺, lin−(CD11c, SiglecF, Ter119, Ly6G) cells) (left) and characterization of eYFP expression (right). a Percentage of eYFP+ monocytes in blood and adventitial macrophages of 45-week-old mice (n = 5; two individual experiments). c Representative immunohistological images and d quantification of eYFP+ macrophages stained for F4/80 (n = 3; three individual experiments). e, f EdU incorporation in adventitial macrophages of Flt3^CreRosa26^eYFP mice (14–16 week old). e Representative immunohistological images and f Quantification of EdU incorporation in eYFP− and eYFP+ F4/80+ macrophages (n = 3; 3 individual experiments). g, h Generation of BM chimeric mice by injection of poly(I:C) into Mx1^CreMyb^flox/flox (BM ablation) followed by transplantation of CD45.1 BM. g Representative flow-cytometry analysis showing expression of CD45.1 and CD45.2 on CD45⁺/lin⁻/CD11b⁺/F4/80⁺ macrophages in the adventitia and h percentage of CD45.1 macrophages after chimerism for 3 and 9 months is shown (n = 3 for 3 months; n = 4 for 9 months; seven individual experiments). i, j Pulse labelling of BM-derived cells in Kit^MCMRosa^GFP mice by tamoxifen feeding for 4 weeks. i Representative immunohistological images of aortas and j percentage of labelled blood monocytes (n = 4, 4 independent experiments) and adventitial macrophages (n = 5, 5 independent experiments). Scale bar as depicted in images. Two-sided t-tests was performed and mean ± SD is shown.