Results of staining, double-staining, or untreated enriched CD34⁺ cells.

(A) CXCR4 expression on enriched CD34⁺ cells stained with antibodies to CD34 FITC and CXCR4 PE. (B) Double-staining of enriched CD34⁺ cells with PE-labeled antibodies to CXCR4 and FITC-labeled antibodies to each of the major integrins: LFA-1, VLA-4, and VLA-5. CXCR4⁺ cells within the CD34⁺population were gated, and the levels of staining for LFA-1, VLA-4, and VLA-5 are shown. The solid histogram indicates negative control staining (CTRL) with isotype control antibody. (C, D) Untreated (CTRL), 100 ng/mL PMA, or EDTA pretreated with cord blood CD34⁺cells were perfused into a parallel plate flow chamber and allowed to settle for 1 minute at 37°C on substrates coated with (C) VCAM-1 or (D) ICAM-1. SDF-1: Plates were coated with (C) VCAM-1 or (D) ICAM-1 in combination with SDF-1. CTRL: Plates were coated with HSA. The data in (A) and (B) are from a representative experiment. The data in (C) and (D) are the average of 3 experiments plus or minus SE. (*IndicatesP < .05).