Prevention of leukocyte migration to inflamed skin with a novel fluorosugar modifier of cutaneous lymphocyte-associated antigen
J. Clin. Invest. Charles J. Dimitroff, et al. 112:1008 doi:10.1172/JCI19220 [Go to this article.]

Figure 3
Effects of 4-F-GlcNAc on effector lymphocyte E-selectin ligand and PSGL-1 expression. Lymphocytic lysates were prepared from inguinal LNs draining DNFB-sensitized skin after day 7 of DNFB sensitization. Lysates (40 μg/lane) were separated on reducing 4–20% SDS-PAGE gradient gels, transferred to PVDF membrane, and blotted with mouse E-selectin–Ig (1 μg/ml) (a) or rabbit anti-sera against mouse PSGL-1 (0.5 μg/ml) (b) and relevant AP-conjugated secondary Ab’s (1:2,000) and AP substrate solution. Negative control blots, E-selectin–Ig staining in the presence in 5 mM EDTA or AP-conjugated secondary Ab’s alone, did not show any signal. Please note the induction of lymphocyte E-selectin ligand principally by the 120- and 220-kDa isoforms of PSGL-1, as well as by a 190-kDa glycoprotein from skin-draining LNs of DNFB-sensitized mice (a). Also, though PSGL-1 levels were relatively unchanged (b), lymphocyte E-selectin ligand induction from skin-draining LNs of DNFB-sensitized FucTIV/VII^–/– mice or of DNFB-sensitized mice treated with 100 mg/kg 4-F-GlcNAc was absent (a).