Extended Data Fig. 6: Endothelial Vegf receptor 2 (Vegfr2) signaling in myocardial infarction.

a, Vegf in blood plasma of controls and 3 days post myocardial infarction (MI) by ELISA (control n=12, MI n=13, two-tailed Welch’s t-test). b-e, Blood leukocyte (b), bone marrow HSPC numbers (c, d), and proliferation (e) obtained in naive Cdh5^CreERT2 and Cdh5^CreERT2;Kdr^fl/fl mice. Both groups were treated with Tamoxifen (B, n=8 mice per group; C-E, n=6 mice per group; two-tailed Mann-Whitney test). f, Experimental outline. Bone marrow mononuclear cells (BMNC) were isolated from Vav1^Cre and Vav1^Cre;Kdr^fl/fl mice and transplanted into lethally irradiated wild type recipients. 8 weeks after bone marrow transplantation (BMT), baseline blood data were obtained by flow cytometry. Both groups were then subjected to MI.g, Blood monocyte and neutrophil numbers 8 weeks post BMT (n=13 Vav1^Cre recipients, n=15 Vav1^Cre;Kdr^fl/fl recipients, two-tailed Welch’s t-test). h, Blood myeloid cell numbers 3 days post MI (n=8 mice per group, two-tailed Welch’s t-test). i, j, Bone marrow Lin⁻ Sca-1⁺ c-kit⁺ (LSK), common myeloid progenitor (CMP) and granulocyte/monocyte progenitor (GMP) numbers (i) (n=8 Vav1^Cre recipients, n=7 Vav1^Cre;Kdr^fl/fl recipients, two-tailed Mann-Whitney test) and proliferation (j) 3 days post MI (n=6 mice per group, two-tailed Mann-Whitney test). k, Image and analysis of colony forming units (CFU) for hematopoietic progenitor cells in blood taken 3 days after MI (n=6 mice pre group, two-tailed Mann-Whitney test). l, Cxcl12 expression in flow-sorted bone marrow endothelial cells on day 2 after MI (n=7 Cdh5^CreERT2, n=10 Cdh5^CreERT2;Kdr^fl/fl, two-tailed Mann-Whitney test). m, Relative expression of Vegfa in flow sorted BMEC isolated from Cdh5^CreERT2 and Cdh5^CreERT2;Kdr^fl/fl mice 3 days post MI by qPCR (n=4 mice per group, two-tailed Mann-Whitney test). n, Vegfa expression in different organs in controls and 48hrs post MI (n=5 mice per group, individual two-tailed Mann-Whitney tests comparing control and MI for each organ). o, mRNA levels of Vegfa (in relation to Gapdh) in FACS-isolated bone marrow cell populations from controls and mice 48hrs after MI (n=5 mice per group, individual two-tailed Mann-Whitney tests comparing control and MI for each cell type). p, Vegfa gene expression on day 2 after myocardial infarction. Relative gene expression of Vegfa in cell populations flow-sorted from leftventricular healthy (Control) and infarct tissue (MI) in relation to Gapdh, as measured by qPCR (n=6 mice per group, two-sided Mann-Whitney test for all).q, Relative Vegfa mRNA assessed by qPCR in bone marrow endothelial cells (BMEC) FACS-isolated from mice with saline or Angiotensin II (HTN) minipumps for 8 weeks (n=5 mice per group, two-tailed Mann-Whitney test). r, Vegfa expression in BMEC from wild type (Control) and Apoe^−/− mice on a Western Diet for 12 weeks (Athero) (n=6 mice per group, two-tailed Mann-Whitney test). s, Flow cytometry of splenic LSK and myeloid cells in Cdh5^CreERT2 controls and Cdh5^CreERT2;Kdr^fl/fl mice 3 days post MI (n=4 mice per group, two-tailed Mann-Whitney test). Data are displayed as mean±SEM.