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. 2006 Dec;169(6):1965–1975. doi: 10.2353/ajpath.2006.051331

Figure 4.

Figure 4

Stx-2 activates p38 and p42/44 MAPKs, instrumental for NF-κB activation. Phosphorylation of p38 (A) and p42/44 (B) MAPKs in podocytes exposed to Stx-2 (50 pmol/L) for 15, 30, 60, and 180 minutes. Cell lysates were analyzed by Western blot using antibodies against the phosphorylated form of each MAPK. The blots were stripped and reprobed with anti-nonphosphorylated p38 or p42/44 antibodies to confirm equal loading of the proteins on the gel. C: Effect of pharmacological inhibition of p38 and p42/44 MAPKs on Stx-2-induced NF-κB-dependent promoter activity. Podocytes were transfected for 3 hours with NF-κB luciferase reporter gene. Then cells were maintained in serum-free medium for 15 hours before exposure to Stx-2 (50 pmol/L) for an additional 6 hours. The p38 inhibitor SB-202190 (20 μmol/L) and the p42/44 inhibitor PD-98059 (10 μmol/L) were added 1 hour before and during stimulation with Stx-2. Relative luciferase activity is expressed as fold stimulation by assuming control as 1. Data are means ± SEM (n = 3 experiments). *P < 0.01 versus control; °P < 0.01 versus Stx-2.