Fig 1: The S100A4/A6/A10/A11 proteins modulate the inflammatory response in KCs via the MAPK signaling pathway. (A) Following 2 h stimulation of KC with appropriate concentrations of S100A4/A6/A10/A11 proteins, cellular RNA and protein were collected for analysis. RT-qPCR results showed that S100A4 and S100A6 protein stimulation significantly upregulated the mRNA levels of various inflammatory cytokines (TNF-α, IL-1β, IL-6, IL-17, IL-12, IFN-γ) and chemokines (CCL2, CCL4). S100A10 protein stimulation significantly upregulated the mRNA levels of TNF-α, IL-17, and IL-12. Similarly, S100A11 protein stimulation significantly upregulated the mRNA levels of TNF-α, IL-6, IL-17, IL-12, IFN-γ, CCL2, and CCL4. (B) WB analysis revealed that S100A4 and S100A11 activated the p38, JNK, and ERK signaling pathways. In contrast, S100A6 and S100A10 predominantly activated the p38 and JNK signaling pathways. Data are presented as mean ± SD (n = 3 independent experiments). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001
Fig 2: S100A4/A6/A10/A11 proteins directly activate HSCs and induce fibrogenic responses. (A) HSCs were individually stimulated with recombinant S100A4/A6/A10/A11 proteins at a concentration of 500 ng/mL for 12 h. Following treatment, RNA and protein samples were collected from the HSCs for analysis. RT-qPCR analysis revealed that all four S100A proteins significantly upregulated the mRNA expression of α-SMA, collagen I, collagen III, Fn1, and MMP1 in HSCs. Regarding TIMP1 mRNA expression, S100A6 upregulated its expression, whereas S100A10 and S100A11 downregulated it. (B) WB analysis demonstrated that stimulation with S100A4/A6/A10/A11 proteins consistently increased the protein levels of α-SMA, collagen I, and collagen III in HSCs. WB also revealed activation of the TGF-β/SMAD2 signaling pathway components. Data are presented as mean ± SD (n = 3 independent experiments). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001
Fig 3: S100A4/A6/A10/A11 proteins activate the TGF-β/SMAD2 pathway in HSCs via KCs. (A) KCs were individually stimulated with recombinant S100A4/A6/A10/A11 proteins. Subsequently, the conditioned media (supernatants) from these stimulated KCs were mixed 1:1 with DMEM and used to treat HSCs for 24 h. Following treatment, RNA and protein samples were collected from the HSCs for downstream analysis. RT-qPCR analysis revealed that conditioned media from KCs stimulated with S100A4 or S100A6 significantly upregulated the mRNA expression of α-SMA, collagen I, collagen III, Fn1, and MMP1 in HSCs, while concurrently downregulating TIMP1 mRNA expression. In contrast, conditioned media from KCs stimulated with S100A10 or S100A11 significantly upregulated the mRNA expression of α-SMA, collagen I, collagen III, and Fn1 in HSCs, while similarly downregulating TIMP1 mRNA expression. (B) WB analysis demonstrated that conditioned media from KCs stimulated with S100A4/A6/A10/A11 proteins collectively upregulated the protein levels of α-SMA, collagen I, and collagen III in HSCs, primarily through the activation of the TGF-β/SMAD2 pathway. Data are presented as mean ± SD (n = 3 independent experiments). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001
Supplier Page from Abcam for Recombinant Mouse S100A10 protein (His tag C-Terminus)