Figure 6.

Fate-mapping aorta intima resident macrophages in the progression of atherosclerosis.

(A) CX3CR1^creER Rosa26-lsl-Tomato Ldlr^−/− mice were treated with tamoxifen by oral gavage three times over a week, then sacrificed and imaged by en face whole mount confocal microscopy. (B) Blood monocyte labeling efficiency following tamoxifen treatment of CX3CR1^creER Rosa26-lsl-Tomato Ldlr^−/− mice, showing Tomato expression as assessed by flow cytometry for classical monocytes (CD115⁺ CD11b⁺ Ly6G⁻ Ly6C⁺) at day 0 (left panel). Labeling kinetics for classical and non-classical monocytes in the blood tracked for 14 days after tamoxifen treatment (right panel). (C) Following tamoxifen treatment, CX3CR1^creER Rosa26-lsl-tomato Ldlr^−/− mice were rested on chow diet for three weeks, then assayed by whole mount en face confocal microscopy for Tomato (red) labeling of Mac^AIR co-stained with CD45 (white). (D) Serum cholesterol levels of CX3CR1^creER Rosa26-lsl-tomato Ldlr^−/− mice following labeling regiment and started on HFD feeding.(E-I) Following tamoxifen treatment, CX3CR1^creER Rosa26-lsl-tomato Ldlr^−/− mice were rested on chow diet for three weeks, then fed HFD for 10-days (E-F), 14 days (G), 28 days (H), and 84 days (I). Mac^AIR were tracked based on expression of Tomato and recruited cells were stained by antibody labeling, as indicated on the micrographs. (J) Following Mac^AIR labeling, the frequency of Tomato+ cells was calculated across the course of disease, from 0-84 days of HFD feeding. Data are derived from (A) n=9 from three independent experiments, (B) n=6 from two independent experiments, (C) n=6 from two independent experiments, (D) n=13 from a single experiment, (E-J) n=26 across all time points from three independent experiments. Error bars are presented as SEM.