Figure 8

JNK signaling plays a critical role in proinflammatory cytokines production. MCPIP1 knockout mice were treated with SP600125 (15 mg/kg, iv) 60 min prior to MCAO and proteins extracted from the ischemic hemisphere of MCPIP1 knockout mice undergoing ischemia 30 min followed by 30 and 90 min reperfusion separately. (A) A representative western blot shows protein levels of JNK phosphorylation. (B) Densitometric analysis was used to quantify p-JNK protein levels versus total JNK in 3 independent western blots and the data are expressed as the normalized folds with respect to sham. Values represent mean ± SD, *p < 0.05 versus sham-treated control. (C) A representative western blot shows protein levels of c-jun phosphorylation. (D) Densitometric analysis was used to quantify p-c-cun protein levels versus total JNK as the loading control in 3 independent western blots and the data are expressed as the normalized folds with respect to sham. Values represent mean ± SD, *p < 0.05 versus sham-treated control. (E) The expression of proinflammatory cytokines was determined at 12 h and 24 h after MCAO and inhibition of JNK activation significantly reduced mRNA levels of TNFα and IL-1β in the ischemic hemisphere of MCPIP1 knockout mice undergoing MCAO. Values represent mean ± SD,* p < 0.05 versus sham group. n = 5 per group.