Fig. 2: Activation of NF-κB and MAPK signaling by CXCL4 and TLR8 in human monocytes.

a, b Immunoblot of whole-cell lysates from monocytes stimulated with CXCL4 (10 μg/ml) and/or TLR8 ligand ORN8L (20 μg/ml) for indicated time course. Data are representative of 4 (a, the samples derive from the same experiment and the gels/blots were processed in parallel) or 2 (b) experiments. Molecular mass markers are displayed to the left of panels. c Flow cytometric analysis of NF-κB p65 phosphorylation in cells pretreated with 1 mg/ml chondroitinase ABC (CHO) and 1 mg/ml heparinase III (H) for 1 h, and then stimulated with CXCL4 for 1 h. Left panel depicts representative histograms and right panel depicts data from 4 experiments. d, e Flow cytometric analysis of NF-κB p65 phosphorylation in monocytes pretreated with 1 µM Bafilomycin A (BafA1) for 30 min, then stimulated with CXCL4 and/or ORN8L for 1 h. d Representative histograms and e cumulative data from 4 experiments. f, g qPCR analysis of mRNA amounts normalized relative to GAPDH mRNA in cells stimulated with CXCL4 and/or ORN8L after treatment with BafA1 (f) or the combined MAPK inhibitors SB 202190 (p38), JNK inhibitor II and U0126 (MEK1/2) used at 10 μM (g). Data in (g) are related to and uses some of the same samples as Supplementary Fig. 5c. Data are from 5 (f) or 4 (g) independent experiments. Data are depicted as mean ± SEM (c, d–g). ****p ≤ 0.0001; **p ≤ 0.01; *p ≤ 0.05 by two-way ANOVA. Source data are provided as a Source data file.