Figure 1. Contribution of DAMPs and DAMP receptors in the promotion of HSC activation and liver fibrosis.

A-B. 2x10⁵ primary quiescent mouse HSCs from BALB/c or αSMA-RFP transgenic mice were co-cultured with increasing amounts of dying primary mouse hepatocytes, followed by fluorescent microscopy, αSMA Western blot or qPCR for Acta2 (n=3 technical replicates). C. Gene expression in primary mouse HSCs (n=4 biological replicates) and untreated whole liver (n=3 biological replicates) from BALB/c mice was compared by RNA-seq; shown are the top 25 enriched receptor genes (left panel). Expression of P2ry14 and other candidate DAMP receptors in mouse HSCs was confirmed by qPCR (n=5 HSC, n=5 livers from the same mice, right panel, biological replicates). D. Primary mouse HSCs from BALB/c mice were treated with recombinant HMGB1, Adora2a agonist CGS 21680, or sphingosine-1-phosphate followed by Western blot for αSMA (n=1 per condition). E-H. AGER knockout (n=10 WT and n=12 KO) (E), TLR4^ΔHSC (n=5 TLR4^fl/fl and n=5 TLR4^ΔHSC) (F), ADORA2A knockout (n=9 WT and n=10 KO) (G), and S1PR3 knockout mice (n=11 WT and n=13 KO) (H) and their respective controls were treated with six injections of CCl₄ (0.5 ml/kg), followed by analysis of the Picrosirius Red area. Data are shown as means ± SEM. Scale bar 200 μm.