Figure 1.

Modulation of c-Rel containing DNA-binding complexes by LPS with or without GlcN. (A) BV2 cells were untreated or pretreated with 5 mM GlcN for 2 h and incubated with 0.1 μg·mL⁻¹ LPS in the presence of or absence of 5 mM GlcN. Nuclear extracts were prepared at 30 min or 12 h and DNA binding to a ³²P-labelled NF-κB probe was measured by EMSA (left panel). Supershift analysis using antibodies targeting p65, p50, c-Rel, RelB or p52 were performed at 15 min or 12 h after LPS stimulation (right panel). Data are representative of at least three independent experiments. (B) RAW264.7 cells were transfected with iNOS promoter-reporter constructs along with the indicated combinations of p65, p50 and c-Rel and stimulated with LPS (0.1 μg·mL⁻¹) with or without 5 mM GlcN as indicated. Luciferase activity was measured at 24 h and shown as relative luciferase activity. ^#P < 0.01 control versus LPS-treated; ^@P < 0.05 control versus LPS-treated; *P < 0.05 LPS versus LPS+GlcN-treated; **P < 0.01 LPS versus LPS+GlcN-treated.