Fig 2: Caspase-11 induction via IL-1/IL-1R is Myd88-dependent in vitro. WT, myd88−/−, trif−/−, and myd88/trif−/− macrophages stimulated with IL-1α, IL-1β, IL-18, HMGB1, LPS, or in combination for 4 h. Induction of caspase-11 gene expression assayed via (A) RT-qPCR via relative copy number and (B) Western with loading control calreticulin (CALR) (representative). Two-way ANOVA using a Tukey's multiple comparison test was performed. **p <0.01, ***p <0.001, ****p <0.0001, n = 3 independent experiments.

Fig 3: Lysozyme induces pain without inflammation. (A–C) Effect of lysozyme (A), LPS (B) and HMGB1 (C) injections on mouse foot paw inflammation. (D–F) Effect of lysozyme (D), LPS (E) and HMGB1 (F) injections on mouse foot paw pain sensitization. (G–I) Effect of lysozyme injections on mice foot paw pain sensitization in (G) C57BL/6J (H) C3H/HeJ (I) B6.129-Tlr2tm1Kir/J. (Figure 1A–I, n=8-16 animals per group, two-way ANOVA, Tukey’s multiple comparison, p<0.001). (J) Effect of lysozyme injections on foot paw inflammation in C57BL/6J, C3H/HeJ and B6.129-Tlr2tm1Kir/J mice (n=12 animals per group, two-way ANOVA, Tukey’s multiple comparison p<0.001). (K–M) Effect of LPS injections on mice foot paw pain sensitization in C57BL/6J (K), C3H/HeJ (L), and B6.129-Tlr2tm1Kir/J mice (M). (Figure 1K–M, n=12 animals per group, two-way ANOVA, Tukey’s multiple comparison, p<0.001) (N) Effect of LPS injections on foot paw inflammation in C57BL/6J, C3H/HeJ and B6.129-Tlr2tm1Kir/J mice (n=12 animals per group, two-way ANOVA, Tukey’s multiple comparison p<0.001). ****p> 0.0001; ns, not significant.