Fig. 5
Treg administration improves the outcome of myocardial infarction. a Tregs sorted by DEREG mice were injected into the border region of the myocardial infarction and visualized as EGFP+ cells in heart section at day 3 after surgery, detecting either the endogenous green fluorescence (upper panel) or using anti-EGFP antibodies (purple, lower panel). Scale bar, 25 μm. b Quantification of infarct size expressed as percentage of the left ventricular area in control (C, circles) and Treg-injected (Treg, squares) mice at 3 months after myocardial infarction. c US imaging analysis of the left ventricular ejection fraction of control (C) and Treg-injected (Treg) mice at 1, 4, and 12 weeks (W) after 40 min (min) of coronary artery ligation followed by vessel reperfusion. d US imaging analysis of the left ventricular ejection fraction of control (C) and Treg-injected (Treg) mice at 1, 4, and 12 weeks (W) after permanent ligation of the left descendent anterior (LAD) coronary artery. e Sections of the heart of Treg-injected mice, showing EdU incorporation (stained red) in the nuclei of CMs (stained green by anti-α-actinin antibodies). Scale bar, 100 μm. f Quantification of the EdU incorporation (% of total CM nuclei) in CMs in control (C) and Treg-injected (Treg) hearts at 15 days after permanent coronary artery ligation. g Sections of the heart of Treg-injected mice, showing EdU incorporation (stained red) in the nuclei of CMs (stained green by anti-PCM1 antibodies). Nuclei are stained in blue with DAPI. Scale bar, 10 μm. h Section of the heart showing injected Tregs (labeled in white by anti-EGFP antibodies) in proximity of one EdU+ CM (EdU is labeled in red and CMs in green by anti-α-actinin antibodies). Nuclei are stained in blue with DAPI. Scale bar, 50 μm. All values are mean ± s.e.m. each dot indicates a biological replicate. Pairwise comparison was performed with the Student’s t-test (b, f). Two-way ANOVA for repeated measurements was used in (c, d). *P < 0.05, **P < 0.01, relative to control
