TLR gene expression by CAR cells and IL-6 levels in reciprocal Tlr4^−/− mice upon chronic repetitive LPS treatment. (A) Experimental outline depicting isolation of CAR cells from BM as well as conventional dendritic cells (cDCs; CD3e^–CD19^–NK1.1^–CD11c^highCD45RA^–MHCII⁺) and plasmacytoid dendritic cells (pDCs; CD3e^–CD19^–NK1.1^–CD11c⁺CD45RA⁺MHCII^high) from spleen of steady-state WT mice for the assessment of Tlr expression as shown in panel B. (B) Tlr1-9 expression as assessed by qRT-PCR in CAR cells (red bars) and DCs (pooled cDCs and pDCs; blue bars) isolated from steady-state WT mice. Expression values were normalized to the housekeeping gene Gapdh. (C) Experimental scheme showing induction of LPS-induced inflammation by a single LPS injection. This schedule is applicable to experimental data depicted in panel D. (D) IL-6 plasma and BM lysate levels in Tlr4^−/− BM reciprocal chimeric mice upon a LPS-induced inflammation (red bars) compared with PBS-injected control mice (blue bars). (E) Graphical scheme depicting experimental outline for chronic repetitive LPS-induced inflammation. This schedule is applicable to experimental data depicted in panel F. (F) IL-6 plasma and BM lysate levels in Tlr4^−/− BM reciprocal chimeric mice upon a LPS-induced inflammation (red bars) compared with PBS-injected control mice (blue bars). (G) IL-6 plasma and BM lysate levels in LeprCre;Tlr4^fl/fl mice upon a LPS-induced inflammation (red bars) compared with PBS-injected control mice (blue bars). (H) IL-6 plasma and BM lysate levels in LeprCre;Myd88^fl/fl mice upon LPS-induced inflammation (red bars) compared with PBS-injected control mice (blue bars). *P < .05, **P < .01, ***P < .001, ****P < .0001. i.p., intraperitoneal; ns, nonsignificant.