Fig. 1: Infection potentiates heat stress-induced inflammatory cell death.

A Schematic illustrating the treatment timeline for (i) heat stress (HS) alone, (ii) HS with bacterial infection, or iii) HS with lipopolysaccharide (LPS) priming. B Real-time analysis of cell death in bone marrow-derived macrophages (BMDMs) challenged with HS for the indicated times. C Representative images of cell death in (B) at 16 hours post-treatment (hpt). D Real-time analysis of cell death in Escherichia coli or Citrobacter rodentium-infected BMDMs with or without subsequent HS (43 °C for 30 min). E Representative images of cell death in (D) at 16 hpt. F Real-time analysis of cell death in LPS-primed BMDMs challenged with HS (43 °C for the indicated times). G Representative images of cell death in (F) at 16 hpt. H Real-time analysis of cell death in LPS-primed wild type (WT), Nlrp3^–/–, Aim2^–/–, Nlrp1b^–/–, Nlrc4^–/–, Mefv^–/–, and Pycard^–/– BMDMs following challenge with HS (43 °C for 30 min). I Real-time analysis of cell death in LPS-primed WT, Nlrc1^–/–, Nlrc2^–/–, Nlrc3^–/–, Nlrc5^–/–, Nlrp6^–/–, and Nlrp12^–/– BMDMs following challenge with HS (43 °C for 30 min). J Real-time analysis of cell death in WT, Zbp1^–/–, and Zbp1^∆Zα2 BMDMs following challenge with HS (43 °C for 1 h). K Real-time analysis of cell death in LPS-primed WT, Zbp1^–/–, and Zbp1^∆Zα2 BMDMs following challenge with HS (43 °C for 30 min). L Real-time analysis of cell death in LPS-primed WT control siRNA (si-Con)-treated, WT Zbp1 siRNA (si-Zbp1)-treated, and Zbp1^–/– BMDMs following challenge with HS (43 °C for 30 min). B Data are shown as mean ± SEM; **** P < 0.0001 (two-tailed t-test; n = 4 from 4 biologically independent samples). C, E, G Images are representative of at least three independent experiments. Scale bar, 50 μm. D, F, H–L Data are shown as mean ± SEM; ****P < 0.0001 (one-way ANOVA with Bonferroni’s multiple comparisons test; n = 4 from 4 biologically independent samples). Exact P values are presented in Supplementary data file 2.