Figure 4

Flow cytometry gating strategy to quantify endothelial progenitor cells (EPCs). Initially, debris, red blood cells and platelets were removed from the analysis based on their forward scatter (FCS) vs. side scatter (SSC) properties (R0 plot 1) and then a gate was set on a CD45 vs. dot plot to contain all CD45⁺ events (R1 plot 2). Next, gate R1 events were displayed on a CD34 vs. SSC dot plot (plot 3) and a second gate (R2) was defined in a sequential strategy to include CD34⁺ events. CD34⁺ cells with low SSC and low CD45 fluorescence (SSC^low/CD45^dim cells) were then gated (R3) (plot 4). The low SSC properties and expression of CD45 were confirmed against total events. Finally, EPCs were identified by a gate (R4) set on a CD34 vs. CD309 (VEGFR2/KDR) dot plot and were defined as CD45^dim/CD309⁺/CD34⁺.