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. Author manuscript; available in PMC: 2011 Mar 8.
Published in final edited form as: J Immunol. 2010 Oct 11;185(10):6205–6213. doi: 10.4049/jimmunol.1000243

FIGURE 4.

FIGURE 4

MAPK/p38 knockdown or MKK3 deletion increases IL-12 production. A, siRNA (10 nM) to p38α or mock siRNA was transfected into BMMφs by nucleofection, and BMMφs were cultured for 48 h. Total RNA was isolated and real-time PCR was performed to analyze p38α mRNA. Insert, Cell lysates from siRNA-transfected macrophages were prepared for Western blotting analysis using Ab to p38α. Total ERK was used as a loading control. B, BMMφs were transfected with mock siRNA for 48 h and treated with saline or SB203580 for 1 h. Parallel BMMφs were transfected with 3, 10, or 30 nM siRNA specific for p38α for 48 h and then stimulated with LPS (10 ng/ml) overnight. Supernatants were harvested to detect IL-12p40 protein by ELISA. C, BMMφs from MKK3–/– mice and control littermates were stimulated with LPS (10 ng/ml) for the indicated times. Cell lysates were prepared for Western blotting analysis using Abs to p-p38, p-MK2, and β-actin as a loading control. D–F, BMMφs from control littermates (filled bars) and MKK3 knockout mice (open bars) were primed with IFN-γ (100 U/ml) and then stimulated with LPS (10 ng/ml) overnight. ELISA was performed to detect cytokine production. Data represent one of three independent experiments (mean ± SD of triplicates for ELISA data). The p values were determined by a Student t test. *p < 0.05.