Detecting Circulating Endothelial Progenitor and Regulatory T Cells

The number of circulating EPCs and Tregs were detected by flow cytometry. The details of flow cytometry were described in our previous paper [20 (link)]. For analysis of EPC cell surface molecules, cells were stained with the following antibodies: unconjugated rabbit anti-CD133 (Abcam, Cambridge, UK), PE-conjugated anti-CD34 (Santa Cruz Biotech, Santa Cruz, CA), VEGFR2⁺ (BD Pharmingen, San Diego, CA) and goat anti-rabbit FITC secondary antibody (Abcam, Cambridge, UK). For analysis of Treg cells, cells were stained with PE-Cy5-conjugated anti CD4 (eBioscience, San Diego, CA) and PE-conjugated anti CD25 (BD Pharmingen, San Diego, CA), and then permeabilized with fixation/permeabiliation working solution and incubated with FITC-conjugated anti-Foxp3 (eBioscience, San Diego, CA).