Flow cytometric quantification of macrophages in the heart and monocytes in the circulation.

<p>(A) Number of cardiac macrophages after PBS perfusion before explanting the hearts. Mean ±SEM; n: Baseline = 10 mice, 3d TAC = 6 mice, 3d Sham = 4 mice, 6d TAC = 8 mice, 6d Sham = 4 mice, 21d TAC = 6 mice, 21d Sham = 5 mice. (B) Number of cardiac macrophages with and without PBS perfusion before explanting the hearts. Mean ±SEM; n of perfused animals: 3d TAC = 6 mice, 6d TAC = 7 mice, 21d TAC = 6 mice; n of non-perfused animals: 3d TAC = 6 mice, 6d TAC = 5 mice, 21d TAC = 6 mice. (C, D) Quantification of cardiac Ly6C^low (C) and Ly6C^high (D) macrophages with and without PBS perfusion before explanting the hearts. Mean ±SEM; n of perfused animals: 3d TAC = 4 mice, 6d TAC = 8 mice, 21d TAC = 4 mice; n of non-perfused animals: 3d TAC = 4 mice, 6d TAC = 7 mice, 21d TAC = 4 mice. (E) Gating strategy to determine the subpopulations of Ly6C^low (black ellipse) and Ly6C^high (blacked dashed ellipse) monocytes in the blood via CD11b and Ly6C staining, previously gating all single cells; concatenated plots of 3 healthy baseline mice. (F, G) Quantification of Ly6C^low (F) and Ly6C^high (G) monocytes in the blood. Mean ±SEM; n: Baseline = 10 mice, 3d TAC = 7 mice, 3d Sham = 5 mice, 6d TAC = 7 mice, 6d Sham = 4 mice, 21d TAC = 5 mice, 21d Sham = 4 mice. (H) Accumulation of Ly6C^high monocyte numbers of all three time points after TAC- and sham-operation. Mean ±SEM; n: Baseline = 10 mice, TAC = 16 mice, sham = 12 mice. *P<0.05, **P<0.01, ***P<0.001.