Fig. 7: CXCL4 and TLR8 signaling crosstalk activates the NLRP3 inflammasome and IL-1β production.

a ELISA of IL-1β protein in culture supernatants (n = 7 independent experiments). b Immunoblot of time course of mature IL-1β protein amounts in culture supernatants (top panel) and whole-cell lysates (bottom panel). c Immunoblot of GSDMD in whole-cell lysates after 6 h stimulation. d Immunoblots of mature Caspase 1 (CASP1) and IL-1β protein in supernatants or whole-cell lysates after 6 h of stimulation with CXCL4 and TLR8 with or without the NLRP3 inhibitor MCC950 (20 μM). Results for 3 different donors are shown. e ELISA of IL-1β protein in culture supernatants after inhibition of CASP1 by AcYVAD (10 μg/ml) (n = 6 independent experiments). f qPCR analysis of NLRP3 mRNA normalized relative to GAPDH mRNA in CXCL4 and/or ORN8L stimulated cells in a time-course experiment (n = 4 independent experiments). g Immunoblots of NLRP3 and AIM2 using whole-cell lysates. h ELISA of IL-1β protein in culture supernatants after 6 h of stimulation with CXCL4 and TLR8 with or without the NLRP3 inhibitor MCC950 20 µM (n = 6 independent experiments). i qPCR analysis of NLRP3 mRNA normalized relative to GAPDH mRNA in CXCL4 and ORN8L co-stimulated cells with/without TBK1/IKKε inhibitor MRT67307 HCl pre-treatment (n = 4). j Immunoblots of NLRP3 and CASP1 using whole-cell lysates. Immunoblot data (b, c, d, g, j) are representative of three independent experiments and data are depicted as mean ± SEM in the other panels. ****p ≤ 0.0001; **p ≤ 0.01; *p ≤ 0.05 by Paired t test, two-tailed (e and h) and two-way ANOVA (f and i). Source data are provided as a Source data file.