Figure 8.

BCL3 controls cytokine production in PMCs without altering the quality of ET. BMMCs were left untreated or incubated with LPS (300 ng/ml) for 24 h to induce ET. Naive and tolerant BMMCs were then stimulated with LPS (1 µg/ml) for 90 min and Bcl3 gene expression was analysed (a). Data show mean ± SD from n = 3 independent experiments. One-sample t-Test was performed to calculate the p-values. Moreover, cells were stimulated with LPS (1 µg/ml) for 4 h and 6 h and lysates were analysed by Western Blotting with antibodies specific for BCL3 and GAPDH (loading control) (b). The Western blot is representative of three independent experiments with three biological samples. Next, BMMCs of WT and Bcl3−/− mice were pretreated with LPS (300 ng/ml) for 24 h to induce ET. Then the cells were stimulated with LPS (1 µg/ml) for 90 min to measure Il6 and Tnf mRNA expression (c), and for 4 h to detect IL-6 and TNF-α protein secretion (d). Also, WT and Bcl3−/− PMCs were incubated with LPS (300 ng/ml) for 24 h to induce ET, and then stimulated with LPS (1 µg/ml) for 4 h to analyse TNF-α protein production (e). Moreover, WT and Bcl3−/− PMCs were stimulated with LPS (1 µg/ml), FSL-1 (1 µg/ml), and IL-33 (2 ng/ml) for 4 h and IL-6 and TNF-α protein secretion was measured (f). Data show mean ± SD from three independent experiments. Student’s t-Test and one-sample t-Test were performed to calculate the p-values. Multiple testing p-values were corrected by FDR. *p < 0.05, **p < 0.01, ***p < 0.001.