Figure 4.
Figure 4. Phenotypic analysis and SDF-1α migration assay. CD34+ cells were obtained via bone marrow aspiration, or following mobilization with AMD3100 or G-CSF from 4 rhesus macaques. CD34+ cells were analyzed by flow cytometry following staining with monoclonal antibodies, including FITC-conjugated anti-human CD49d (VLA-4), PE-conjugated anti-human CD34, and PE-Cy5-conjugated anti-human CXCR4. (A) CXCR4 cell-surface marker expression on CD34+ cells in a representative experiment (animal RQ3636) is shown in the top panel, and a summary of CXCR4 expression on CD34+ cells in all 4 rhesus macaques is shown in the bottom panel. (B) SDF-1α migration assay. CD34+ cells from 4 rhesus macaques were incubated in transwells in the presence or absence of SDF-1α. The percentage migration of CD34+ cells toward SDF-1α in vitro is summarized for each cell population analyzed (BM, AMD3100-, and G-CSF-mobilized cells). (C) VLA-4 cell-surface marker expression on CD34+ cells in a representative experiment (animal RQ3636) is shown in the top panel, and a summary of VLA-4 expression on CD34+ cells in all 4 rhesus macaques is shown in the bottom panel. SD indicates standard deviation.

Phenotypic analysis and SDF-1α migration assay. CD34+ cells were obtained via bone marrow aspiration, or following mobilization with AMD3100 or G-CSF from 4 rhesus macaques. CD34+ cells were analyzed by flow cytometry following staining with monoclonal antibodies, including FITC-conjugated anti-human CD49d (VLA-4), PE-conjugated anti-human CD34, and PE-Cy5-conjugated anti-human CXCR4. (A) CXCR4 cell-surface marker expression on CD34+ cells in a representative experiment (animal RQ3636) is shown in the top panel, and a summary of CXCR4 expression on CD34+ cells in all 4 rhesus macaques is shown in the bottom panel. (B) SDF-1α migration assay. CD34+ cells from 4 rhesus macaques were incubated in transwells in the presence or absence of SDF-1α. The percentage migration of CD34+ cells toward SDF-1α in vitro is summarized for each cell population analyzed (BM, AMD3100-, and G-CSF-mobilized cells). (C) VLA-4 cell-surface marker expression on CD34+ cells in a representative experiment (animal RQ3636) is shown in the top panel, and a summary of VLA-4 expression on CD34+ cells in all 4 rhesus macaques is shown in the bottom panel. SD indicates standard deviation.

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