Fig. 5

Endothelial Dll1 regulates macrophage maturation and arteriogenesis. a Analysis of cell populations by flow cytometry in ischemic muscle of induced endothelial Dll1 mutant mice (Dll1 ^iΔEC) and control mice. n = 8 mice/group, error bars represent s.e.m. *p < 0.05 by two way ANOVA with Bonferroni multiple comparison test. b Gene expression analysis of macrophages sorted from d3 ischemic muscle. n = 3 independent experiments, error bars represent s.e.m. **p < 0.01, *p < 0.05, Student’s unpaired t-test. c–f Rescue of endothelial Dll1 deficiency after HLI by transfer of ischemic macrophages. On d1 after HLI, indicated groups received intramuscular limb injections of 5 × 10e5 macrophages isolated from d3 ischemic muscle (MF) or Ly6C^hi monocytes isolated from spleen (Mo) of wt mice, or received control treatment. c Foot perfusion by laser Doppler perfusion measurement immediately after induction of HLI (0) and during follow up. n = 5/7/7/7 mice/group, error bars represent s.e.m. ***p < 0.001 by two way ANOVA and Bonferroni post test. d Quantification of inner circumference and wall area, and representative H&E stained images of collateral arteries d14. Scale bar=50 μm. n = 7 mice/group, error bars represent s.e.m. ***p < 0.001 vs control by one way ANOVA and Dunnett’s multiple comparison test. e Representative SMA-stained immunofluorescence of muscle sections, Scale bar 100 μm (left) and quantification of arterial branches (SMA⁺) per section d14, n = 5 mice/group, error bars represent s.e.m. ***p < 0.001 vs control by one way ANOVA and Dunnett’s multiple comparison test. f Representative H&E stained muscle sections. Scale bar=50 μm (left) and quantification of ghost cells and regenerating muscle fibers (right) at d14, n = 5 mice/group, error bars represent s.e.m. ***p < 0.001 vs control by one way ANOVA and Dunnett’s multiple comparison test