Fig. 3. TRIF and CD14 mediate TNF-induced lethality.

(A, B) Mouse body temperature (A) and survival (B) over time in B6, Tnfr1^−/−, Tnf^−/−, Trif^−/−, Cd14^−/−, Trif^−/−Cd14^−/− Tlr3^−/−, Tlr4^−/− and Tlr3^−/−Tlr4^−/− mice after intravenous injection with 9ug mTNF. (C-G) Serum levels of (C) TNF, (D) IFNβ, (E) IL-1β, (F) relative lactate dehydrogenase (LDH) and (G) Alanine Aminotransferase (ALT) in indicated mice 4 hours after i.v. injection with 9ug mTNF. Data points indicate individual mice tested (B6 PBS, n=13; Tnfr1^−/− PBS, n=3; Tnf^−/− PBS, n=3; Trif^−/− PBS, n=10; Cd14^−/− PBS, n=7; Trif^−/−Cd14^−/− PBS, n=3; Tlr4^−/− PBS, n=6; Tlr3^−/− PBS; n=9; Tlr3^−/−Tlr4^−/− PBS, n=3 B6 mTNF, n=22; Tnfr1^−/− TNF, n=5; Tnf^−/− TNF, n=5, Trif^−/− mTNF, n=14; Cd14^−/− mTNF, n=10; Trif^−/−Cd14^−/− TNF, n=5; Tlr4^−/− mTNF, n=9; Tlr3^−/− mTNF, n=9; Tlr3^−/−Tlr4^−/− TNF, n=5 ). (H, I) H&E staining of (H) ileum and (I) liver sections from B6 and Trif^−/− mice 4 hours after injection with 9ug mTNF or PBS. Dotted boxes indicate areas of leukocyte recruitment. (J) Representative image of spleens from B6 and Trif^−/− mice injected with 9ug TNF or PBS 6 hours after injection. (K) TUNEL staining and (L) quantification of TUNEL+cells/mm² in the spleen of B6 and Trif^−/− mice 4 hours after injection with 9ug mTNF. Each point represents 1 field of view across 3 biological replicates. Analysis of variance (ANOVA) was used for comparison between groups: ns, nonsignificant (p>0.05); *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.