Figure 2

IRAK4 mutation impacts innate and adaptive cytokine response in MRL mice. Splenocytes from 3 (A), 6 (B), and 10 month old mice (C) were simulated with vehicle (Veh.), E. coli LPS (100 ng/ml), CpG (1 µM) for 18 h (left panels) or with PMA and ionomycin (P:I) for 18 h (right panels). Culture supernatants were obtained at 18 h and assayed for RANTES/-CCL5 (left panels) and (IFNγ) by ELISA. Splenocytes from 3, 6, and 10 month old mice were stimulated with PMA and ionomycin (PMA:I) for 5 h in the presence of Brefeldin A (BFA). Cells were analyzed by flow cytometry, Percentage of intracellular IFNγ expressing cells is shown for CD4 T cells (D) identified as live CD3⁺, CD4⁺ and CD8 T cells (E) identified as live, CD3⁺, CD8⁺. Each dot represents a mouse. Data shown are individual biological replicates +/− s.e.m. Statistical significance was evaluated by one way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 for comparison of individual biological within a treatment group and ⁺p < 0.05, ⁺⁺p < 0.01, ⁺⁺⁺p < 0.001, ⁺⁺⁺⁺p < 0.0001 for comparison of individual biological with the same mouse train in vehicle treated group.