A, Disturbed flow was induced in the LCA of ApoE−/− mice by partial ligation surgery, while the contralateral RCA was used as an internal control. B, Ligated mice were fed a HFD for 4 to 28 days. Shown is a representative micrograph displaying diffuse atherosclerosis (yellow arrows) in LCA, but not in RCA, two weeks post-ligation. C, LCAs (dashed red box) and RCAs (dashed blue box in (B) obtained from 3 mice at a given time point were pooled (regarded as n=1) and vascular leukocytes extracted. D, Shows the gating strategy used for flow cytometry analyses for a representative arterial leukocyte sample harvested from LCA 7 days post-ligation. Ultra-bright counting beads (D2) were used to calculate absolute cell counts. Gate numbers indicate percent of parent. FSC-H, forward scatter height; FSC-A, forward scatter area; SSC-A, side scatter area.