Fig. 1: Synergistic activation of inflammatory genes by CXCL4 and TLR8.

Human blood monocytes were stimulated with CXCL4 (10 μg/ml) and/or TLR8 ligand ORN8L (20 μg/ml). a RNAseq data analysis. K-means clustering (K = 5) of differentially expressed genes induced more than twofold with FDR < 0.05 by CXCL4, ORN8L, and CXCL4 + ORN8L at 6 h (n = 3 independent blood donors). b Gene Ontogeny (GO) analysis of genes upregulated by CXCL4 + ORN8L costimulation versus resting cells. c mRNA of IL6, IL12B, and TNF was measured by quantitative PCR (qPCR) and normalized relative to GAPDH mRNA. Cumulative data from 4 experiments. d IL6 and TNF protein levels in the supernatant of cultured cells were measured by ELISA. Cumulative data from 5 experiments. e Heat map depicting expression of representative cytokine and chemokines from (a) presented relative to maximum expression. f Nanoparticle formation measured using dynamic light scattering. Number mean = the average size of nanoparticles. ND = not detected. The number mean is from one sample measured in triplicate and is representative data out of 3 experiments. g Flow cytometric analysis of the internalization of ORN8L-AF488 after 30 min incubation in the absence or presence of CXCL4 in human monocytes. Left panel, representative FACS plot; right panel, cumulative data from 3 experiments. Data was depicted as mean ± SEM; ****p ≤ 0.0001; ***p ≤ 0.001; **p ≤ 0.01; *p ≤ 0.05 by two-way ANOVA (c) and one-way ANOVA (d, g). Source data are provided as a Source data file.