In Vivo Ischemia and Wound Healing

All animal experiments were approved by Emory University IACUC. Hindlimb ischemia^{32 (link)} and skin wound^{33 (link)} were performed on male athymic nude mice as we described previously. For the hindlimb ischemia studies, the estimated minimal number for having a meaningful difference at 4 weeks is 5, so we included 5 to 10 animals in each group (n = 6 for PBS-, 10 for HUVEC-, 5 for rEC-injected groups). Before sacrifice for tissue harvest, the mice were perfused with fluorescein-labeled Griffonia (Bandeiraea) Simplicifolia Lectin I (BSL1, Vector Laboratory Inc.) by direct cardiac injection to stain functional endothelial cells in blood vessels. The tissue sections were processed for confocal imaging with a Zeiss LSM 510 Meta confocal laser scanning microscope and LSM 510 Image software (Carl Zeiss).
Details on the materials and methods, including the following items, can be found in the online-only Data Supplement: Flow cytometry^{32 (link)}; Acetylated-LDL uptake and UEA1 lectin staining^{32 (link)}; In vitro tube formation assay^{32 (link)}; Immunohistochemistry and immunocytochemistry^{32 (link)}; Real-time RT-PCR (Table 1 in the online-only Data Supplement)^{32 (link)}; Microarray; Heat map and clustering analysis^{34 (link),35 (link)}; RNA-seq analysis; Statistical analysis.