Genomic deletion of STGal-4. (A) Schematic of vector used to create knockout cells. ST3Gal-4 genomic DNA sequencing results for WT HL-60s (top row) and the 2 alleles in the ST3Gal-4-KO cells (middle and bottom row). Here, the solid box highlights the guide RNA sequence and the dashed box denotes the protospacer adjacent motif (PAM/NGG). Seventeen- and 238-bp chromosomal deletion are noted upon genomic editing. (B) Flow cytometry histograms measuring CLA/mAb HECA-452, sLe^X/CSLEX-1, Le^X/HI98, and ECL lectin binding. Solid empty histograms correspond to WT cells; dashed histograms correspond to isotype control except for last panel where it is secondary Ab alone; and gray shaded histograms are for ST3Gal-4-KO HL-60s. (C-F) WT and ST3Gal-4-KO cell rolling data on IL-1β–stimulated HUVECs (C-D), recombinant P-selectin-IgG (E), and L-selectin-IgG (F) at 1 dyne/cm². Statistics symbols are identical to Figure 3. P2H3 is an anti-E-selectin blocking mAb. ST3Gal-4-KOs did not roll on L- and P-selectin. Residual cells that rolled on E-selectin displayed fivefold higher rolling velocity compared with WT HL-60s.