Neutrophils derived from hHSCs. hHSCs were differentiated to mature neutrophils over 12 days. On the fifth day of the differentiation process, the cells were transduced with lentivirus containing either ST3Gal-4 shRNA (“ST3Gal-4⁻”) or TRC control virus (“DsRed”). In both cases, DsRed serves as a reporter of transduction. (A) CD34 and CD11b expression changes with time confirming differentiation to granulocytes. (B) Transduction efficiency quantified based on the percentage of cells that are DsRed positive. (C-E) Rolling and adherent cell density data for neutrophils derived from hHSCs on IL-1β–stimulated HUVECs (C), recombinant P-selectin (D), and recombinant L-selectin (E). Fluorescence microscopy was applied in panels C-E, so that only DsRed-positive cells were analyzed in the flow chamber assay. Knocking down ST3Gal-4 reduced cell adhesion to all 3 selectins. Data are presented for hHSCs derived from 4 different human donors. Statistics symbols are the same as Figure 3.