Fig. 6.

Blockade of p38 MAPK pathway inhibits O-GlcNAcylation and activation of NF-κB/p65 in LPS-treated endothelial cells. BAEC were pretreated either with SB203580 (10 μM, a specific inhibitor of p38 MAPK) or PD98059 (10 μM, a selective inhibitor of MAPK kinase or MEK) for 1 h followed by LPS (100 ng/ml) incubation for 1 h. The treated cells were subjected to (A) Western blotting of proteins either from WGA-enriched O-GlcNAc modified proteins or whole cell lysates (to assess protein input as a loading control); (B) immunofluorescent staining of NF-κB or DAPI with a kit including ProLong® Gold and SlowFade® Gold Antifade. The secondary antibody was conjugated with a fluorescent green dye (Alexa Fluor 488). Nuclear p65 fluorescence was quantified. All images shown are representative of three independent experiments. Data are expressed as means ± SEM (n=3). ^#P<0.05, ^##P<0.01 compared to the vehicle-treated control group; *P<0.05, **P<0.01 compared to the LPS-only group.