Figure 1: CARD9 negatively regulates IL-1β production in vitro and in vivo.

(a,d,g) IL-1β secretion (as measured by ELISA), (b,e,h) cellular viability (as measured by LDH release) and (c,f,i) TNF-α (as measured by ELISA) of WT, Nlrc4^−/− and Card9^−/− BMDMs after infection with S. Typhimurium SL1344 at m.o.i.’s 1, 10 and 50 for 2 (a–c), 6 (d–f) and 24 (g–i) h. (j,m) IL-1β secretion, (k,n) cellular viability and (l,o) TNF-α after infection of WT and Card9^−/− BMDMs with E. coli P19A at m.o.i.’s 1, 10 and 50 for 2 (j–l) and 6 (m–o) h. (p) Immunoblot analysis of pro-IL-1β, caspase-1 and β-actin in spleen cells isolated from infected WT and Card9^−/− C57BL/6 mice after intravenous infection with S. Typhimurium M525P (4 × 10³ colony-forming units). (q) Densitometric analysis of this immunoblot. *P<0.05 (one-way analysis of variance with Tukey’s multiple comparisons test). (a–i) Data from five independent experiments (mean and s.e.m.). (j–o,q) Data from two independent experiments (mean and s.e.m). (p) Representative data from two independent experiments, using cells pooled from four to six mice per genotype, plus two negative controls per genotype. Mice were from 8 to 16 weeks old, both male and female.