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. Author manuscript; available in PMC: 2022 Oct 29.
Published in final edited form as: Nat Immunol. 2022 Apr 29;23(5):705–717. doi: 10.1038/s41590-022-01192-4

Figure 2. Low cytosolic concentration of LPS triggers noncanonical activation of the NLRP3 inflammasome when cytosolic bacterial mRNA is also present.

Figure 2.

a-b, Immunoblots of macrophage concentrated supernatants (20hr) or WCE (6hr), and cytokine concentrations and LDH release in culture supernatants (20hr) as indicated following transfection of ultrapure LPS (low dose 2ng/mL or high dose 1μg/mL) +/− mRNA from E. coliLPSmut or in vitro transcribed (IVT) (100ng/mL), or transfection of LPS (2ng/mL) +/− mRNA from E. coli or eukaryotic cells (100ng/mL) (a), and transfection of wild-type (WT) or Nlrp3–/– macrophages with indicated doses of ultrapure LPS or mRNA from E. coliLPSmut, L. innocua, or IVT (b). Bacteria:macrophage ratio=20:1, except 50:1 for F. novicida. LDH measured by cytotoxicity assay; IL-1β, TNF and IL-6 by ELISA. Error bars, mean ± s.e.m. One-way ANOVA followed by multiple comparisons Sidak tests and p values indicated in bar graphs in a (IL-1β: n=4, LDH: n=3), and b (n=4). Cleaved caspase-11 corresponds to caspase-11 p30. Results represent at least 3 independent experiments.