Multiparametric fluorescence activating cell sorting analysis was used to assess mobilization and demargination of circulating monocytes. A: representative dot plots from lean and DIO mice showing the gating strategy used to identify the circulating live monocyte subpopulations: total (CD115+), inflammatory (CD11c−, Ly6CHi), or resident/patrolling (CD11c−, Ly6CLow) monocytes. B–D: quantification of the percent circulating live total, inflammatory, and resident/patrolling monocytes in peripheral blood at baseline (Base) or 1 or 3 days postligation. B: there were significantly more total monocytes at baseline in DIO versus lean mice. The total monocyte percentage significantly increased from baseline to day 1 in both groups (mobilization) and then decreased from days 1 to 3 in both groups (demargination). C and D: the significant changes observed in the total monocyte population were mirrored in the Ly6CLow/patrolling monocyte population but not in the Ly6CHi/inflammatory monocyte subset.