Fig 1: Serum S100A8/A9 concentrations between groups. The boxplots illustrate medians, quartalies and ranges of serum S100A8/A9 concentrations between groups [µg/ml]. a SLE patients versus healthy controls. b–d NPSLE patients versus non-NPSLE patients according to three NPSLE attribution models (b ACR model, c SLICC A model, d SLICC B model)
Fig 2: Induction of S100 proteins by hyperosmolarity in different ocular surface epithelial cells. Cells were exposed to a hyperosmolar medium (70 mM NaCl [orange] or 90 mM NaCl [green]) for 24 h and (A) analyzed for mRNA expressions of S100A4, S100A8, and S100A9 via RT-qPCR. (B) After 48 h of hyperosmolarity exposition, the protein releases of S100A4, S100A8, S100A9 and S100A8/A9 were analyzed via ELISA in a Wong–Kilbourne derivative of the Chang conjunctival (WKD) cell line. (C) The mRNA expression of alarmins were analyzed (D) and protein release in a human corneal epithelial (HCE) cell line. The experiment was repeated four times (n = 4). Results are expressed as a fold change between treated and non-treated (NT) conditions and were analyzed via ANOVA statistical test (Kruskal–Wallis), followed by Dunn’s post hoc test: * p = 0.05, ** p = 0.01 and *** p = 0.001. The horizontal line corresponds to the value of the non-treated cells reported as 1 (n = 4).
Fig 3: Action of DAMPs on the epithelial cell release of cytokines. Epithelial cells were exposed to a normal medium with protein recombinant (S100A4 [pink], S100A8 [purple], or S1009 [blue] at 0.1 and 1 µg/mL or HMGB1 [green] at 100 or 300 ng/mL) for 48 h and then analyzed for protein release of IL6, IL8, TNFa, and MCP1 via ELLA and Multiplex in (A) a Wong–Kilbourne derivative of the Chang conjunctival cell (WKD) line and (B) a human corneal epithelial (HCE) cell line. The experiment was repeated four times (n = 4). Results are expressed as a fold change between treated and non-treated (NT) conditions and were analyzed via ANOVA statistical test (Kruskal–Wallis), followed by Dunn’s post hoc test. The horizontal line corresponds to the value of the non-treated cells reported as 1.
Fig 4: Effects of DAMPs and MCP1 in macrophage migration. In the CytoSlectTM 24-well cell migration assay, THP1 was differentiated for 48 h. (A) Next, macrophages were exposed to S100A4 (2 ng/mL [pink]), S100A8 (3 ng/mL [purple]), S100A9 (2 ng/mL [blue]), HMGB1 (100 ng/mL [green]), or MCP1 (100 ng/mL [yellow]). (B) Macrophages were exposed to the same DAMPs with or without RAGE inhibitors (RAP [angled hatches]) and TLR4 (TAK [horizontal hatches]). The experiment was repeated three times (n = 3). The quantification of cells in the top and bottom of the insert was measured via fluorescence. Results are expressed as a fold change between treated and non-treated (NT) conditions and were analyzed via ANOVA statistical test (Kruskal–Wallis), followed by Dunn’s post hoc test. * p = 0.05. The horizontal line corresponds to the value of the non-treated cells reported as 1.
Supplier Page from Thermo Fisher Scientific for Human Calprotectin L1/S100-A8/A9 Complex ELISA Kit