Figure 1.
IL-1β cytokine production in PBMCs after 24h stimulations. Cells were stimulated with MSU crystals (300 μg/mL), palmitate (50 μM) and the combination of both. Concentration of IL-1β were measured in supernatants by ELISA. The first cohort was split into (a) 181 normouricemic controls and (b) 123 gout patients. Similar analysis was performed in other cohorts which consist of (c) 132 healthy individuals and (d) 148 gout patients. Group differences were assessed by paired Wilcoxon test and the p values are reported.
Figure 2.
RNA-seq after 24h PBMC stimulation with MSU crystals.
(a) Principal Component Analysis Biplots of the 3 experiments. The first experiment contains 2 replicates (gout patients, n=4 and n=3), where cells were stimulated with MSU crystals (300 μg/mL), palmitate (50 μM), palmitate+MSU, LPS (10 ng/mL) or LPS+MSU. The second experiment (healthy individuals, n=4) was independently performed in the same experimental conditions with MSU, palmitate and palmitate+MSU stimulations. Clustering along the PC1 can be observed based on the experimental conditions (MSU with Control, C16 with C16+MSU, LPS with LPS+MSU) showing similar transcriptomic signatures associated with each pair. (b) Volcano plots depicting the results of the Differential Expression Analysis comparing cells stimulated with MSU crystals and control. X axis represents the log2 Fold Change and y axis the p value for each analyzed mRNA.
Figure 3.
RNA-seq after 24h PBMC stimulations with palmitate, LPS and MSU combinations. The first experiment contains 2 replicates (gout patients, n=4 and n=3), where cells were stimulated with MSU crystals (300 μg/mL), palmitate (50 μM), palmitate+MSU, LPS (10 ng/mL) or LPS+MSU. The second experiment (healthy individuals, n=4) was independently performed in the same experimental conditions with MSU, palmitate and palmitate+MSU stimulations. Volcano plots depicting the results of the Differential Expression Analysis comparing (a) palmitate vs control; (b) palmitate+MSU vs palmitate; (c) LPS vs RPMI and (d) LPS+MSU vs RPMI. X axis represents the log2 Fold Change and y axis the p value for each analyzed mRNA. (e) KEGG pathway enrichment analysis of the top upregulated pathways in all comparisons. Infectious disease terms were excluded.