Illustration of FACS gating analysis of angiogenic cells (AC133⁺/KDR⁺PBMCs).

Representative flow cytometric density plots demonstrating the gating protocol used to identify angiogenic cells: A) PBMCs stained with PE-conjugated and APC-conjugated mouse IgG (isotype controls) for non-specific fluorescent signals; B) PBMCs stained with antibodies against human CD133 (AC133) (PE-conjugated) and VEGFR-2 (KDR) (APC-conjugated) for AC133⁺/KDR⁺PBMCs; A1, B1) FSC-SSC density dot plots of Ficoll-isolated PBMCs and R1 was gated for monocytes; A2, B2) FL2 (PE)-FL4 (APC) density dot plots of R1-gated monocytes. Angiogenic cells = cells in B2 upper-right quadrant – cells in A2 upper-right quadrant.