Figure 2. Signal transduction, gene expression, and cytokine release induced by TNF in Tnfrsf1a^+/– mice.

(A) Quantification of NF-κB EMSA results in nuclear extracts of BMDMs after stimulation with 10 ng/ml TNF. The graph shows intermediate activation of NF-κB in the Tnfrsf1a^+/– samples compared with the Tnfrsf1a^+/+ and Tnfrsf1a^–/– samples. (B and C) Quantification of Western blot results showing JNK and ERK phosphorylation in BMDMs at several time points after stimulation with 10 ng/ml TNF. See also Supplemental Figure 2, B–D, for original blots. (D–F) Intermediate induction of genes coding for IKBA, IL-1β, and IFN-β in livers of mice injected i.p. with 30 μg TNF. Data for Tnfrsf1a^–/– mice are not shown, because these mice are not responsive to TNF. (G–I) Intermediate IL-1β, IL-12p40, and NO_x release in sera of Tnfrsf1a^+/– mice injected with 30 μg TNF. (J and K) Cumulative protection against hypothermia and lethality induced by injection of 37.5 μg TNF (1.5-times LD₁₀₀ in Tnfrsf1a^+/+ mice). Ifnar1^+/+ mice (n = 7) were compared with Ifnar1^–/– mice (n = 10), which were protected by the ROS inhibitor Tempol (n = 9) or by Tempol and IL-1RA (Anakinra) (n = 10). Numbers in parentheses indicate the number of mice in each group that did not survive. Data represent mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 (Student’s t test in D–I and log-rank test in K).