A, The number of colonies for 100 transduced cells from the indicated populations at 10 days of a CFC assay for control and S1PR3OE lentiviral knockdown vectors (n=3). B, Normalized colony distribution showing burst-forming units (BFU) are significantly decreased with S1PR3OE and macrophage (M) colonies are significantly increased from all scored cell populations (n=3 CB). G (granulocyte); GEMM (granulocyte erythrocyte monocyte megakaryocyte). C, Myeloid (CD33+) vs erythroid (GlyA+) lineage distribution for all pooled cells from one representation assay at day 14 from (A).
D, Lineage distribution outcomes in single cell assays performed on MS5 stroma in erythroid-myeloid cytokine conditions. Data for indicated number of cells scored in each condition pooled from 3 CBs. E-F, Average number of GlyA+ or CD33+ cells from single cell differentiation assays (D). G, Human CD45 chimerism, H, Log2 fold change (FC) in BFP which marks transduced cells relative to input, I, CD19+ B lymphoid cells in the transduced fraction and J, CD33+ myeloid cells in BFP+ cells for control or S1PR3OE measured in the injected femur at 4 weeks post-transplant in xenotransplantation, 5 mice for each condition from 3 CB. *** P<0.001** p<0.01, *<0.05, unpaired Student t-test. Data are mean and s.d. except for E-F where data are mean and s.e.m.