Figure 6.

Cardiac macrophages (mϕs) regulate angiogenesis. A, Taqman quantitative polymerase chain reaction for Vegfa on cDNA from freshly isolated mϕs from sham or transverse aortic constriction (TAC) operated mice (n=6, 7). Statistical significance by cytokine was determined by 2-tailed unpaired Student t test. B, Representative images (left) of isolectin B4-stained sections (green) derived from sham or TAC-operated mice after isotype or α-CD115 antibody administration. Bar graph (right) shows quantification of microvascular density (vessels/cardiomyocyte; n=5, 7, 12, 12). Statistical significance was evaluated by a 2-way ANOVA. All pairwise comparisons were made. Tukey tests were used to correct for multiple comparisons. C, Representative images of tube formation assay using HUVECs (human umbilical vein endothelial cells) cocultured with negative control (CTL) or with cardiac mϕs compared to positive control. Bar graphs show the quantification of various aspects of tube formation (mesh area, number of nodes, junctions, and meshes; n=3, 6, 3). Statistical significance was evaluated by a Kruskal-Wallis test. All comparisons were made against the negative CTL. Dunn tests were used to correct for multiple comparisons. Data are presented as mean±SEM or median±95% CI. Scale bar=100 µm for C and 200 µm for B. AU indicates arbitrary units; DAPI, 4',6-diamidino-2-phenylindole; and MVD, microvascular density. Statistical significance is summarized as ns, not significant, *P<0.05, **P<0.01, and ***P<0.001.