Figure 4

Expression of angiogenin in migrated and sorted bone marrow cell populations. (A) Immunoreactive angiogenin levels in conditioned media from unfractioned, stromal cell-derived factor 1 non migrated (SDF-1^non) and migrated (SDF-1^mig) mononuclear cells (MNCs; n = 7 per group). (B) Angiogenin mRNA levels of unfractioned, SDF-1^non and SDF-1^mig MNCs (n = 4 per group). Values are means ± standard error of the mean. *P < 0.05, **P < 0.01, ***P < 0.001, versus unfractioned; ^# P < 0.05, ^### P < 0.001, versus SDF-1^non. (C) Angiogenin mRNA levels in sorted bone marrow cells. (i-iii) Gating strategy to sort BM monocyte subpopulations: human MNCs isolated by Ficoll gradient from BM were analyzed by flow cytometry (i). Lympho-monocyte population was gated and analyzed according to CD3 positivity (ii) in order to exclude lymphocytes. Cells negative for CD3 were gated and further analyzed for monocytes markers CD14 and CD16 (iii). Four distinct populations were sorted: CD14⁻/CD16⁺ (pink gate), CD14⁺/CD16⁺ (green gate), CD14⁺/CD16⁻ (red gate), negative cells (black gate). Angiogenin levels were measured in triplicate by RT-PCR (iv). Values are means ± standard error of the mean. **P < 0.01, versus CD3⁻/CD14⁻/CD16⁻ cells. FSC, forward-scattered light; SSC, side-scattered light; UBC, Ubiquitin C.