Fig 1: Gene expression, protein expression, and endothelial tube forming assays. A, Expression of three potent proangiogenic genes in CD34+CD146+, CD34+CD146-, and CD34+ UF ASCs was compared by PCR. CD34+CD146+ ASCs expressed significantly more VEGF (***P < .001) and ANGTP1 (***P < .001) than CD34+CD146- and CD34+ UF ASCs, and significantly greater levels of FGF than CD34+CD146- ASCs (**P < .01). B, Representative immunofluorescence images (top row) and binarized images from ImageJ analysis (bottom row) showing results from endothelial tube forming assays. Human microvascular endothelial cells (green due to staining with Calcein AM) were cocultured for 18 hours in transwells with CD34+CD46+ ASCs (left), CD34+CD146- ASCs (middle), and CD34+ UF ASCs (right). Scale bars = 100 µm. C, Graphs showing results from endothelial tube forming assay analysis demonstrating, C(i), greater total pixel density, C(ii), tube length, and, C(iii), total branching length in HMECs cocultured with CD34+CD146+ compared to CD34+CD146- ASCs (all *P = .05). D, Graphs showing the results from enzyme-linked immunosorbent assays comparing protein expression between three ASC subpopulations. CD34+CD146+ ASCs expressed significantly more VEGF (****P < .0001) (D[i]) and ANGTP1 (****P < .0001) (D[ii]) than both CD34+CD146- and CD34+ UF ASCs, as well as significantly greater levels of FGF than CD34+CD146- ASCs (***P < .001) and CD34+ UF ASCs (****P < .0001) (D[iii]). ANGPT1, angiopoietin-1; ASC, adipose-derived stromal cell; FGF, fibroblast growth factor-2; HMEC, human microvascular endothelial cell; PCR, polymerase chain reaction; UF, unfractionated; VEFG, vascular endothelial growth factor