Aging-related Tet2 ^ΔMye -driven clonal hematopoiesis exacerbates liver fibrosis, and IL-1β–NLRP3 pathway is not involved in the liver fibrosis in Mye-KO-CCl ₄ mice. (A) Chart for the construction of an aging-related Tet2^ΔMye-induced clonal hematopoiesis mouse model with liver fibrosis. BDL was performed to construct a liver fibrosis model in young-BMT and old-BMT mice 6 wk after BMT. (B) Frequency of MDMs in livers of young-BMT and old-BMT mice after BDL for 4 wk (n = 4 for each group). (C) The serum levels of CCL2, CCL8, and IL-6 in young-BMT and old-BMT mice after BDL for 4 wk (n = 4 for each group). (D and E) Picrosirius red staining (D) (scale bar, 125 μm) and quantitative analysis of collagen deposition (E) in livers from young-BMT and old-BMT mice following BDL, treated with Bindarit plus anti–IL-6 Abs (n = 4 for each group). (F and G) Changes of serum Col IV (F) and HA (G) levels in young-BMT and old-BMT mice treated with Bindarit, anti–IL-6 Abs, or Bindarit plus anti–IL-6 Abs (n = 4 for each group). (H) IF (scale bar, 50 μm) and statistical analysis of NLRP3 in the liver of Tet2^WT-CCl₄ and Tet2^ΔMye-CCl₄ mice (n = 5 for each group). Red: anti-NLRP3; blue: DAPI. (I) IHC staining (scale bar, 50 μm) and statistical analysis of NLRP3 in the liver of Tet2^WT-CCl₄ and Tet2^ΔMye-CCl₄ mice (n = 5 for each group). (J) Inhibition of IL-1β in the serum of oil- or CCl₄-treated Tet2^WT and Tet2^ΔMye mice (n = 5 for each group). (K and L) Change of Ccl2 (K) and Ccl8 (L) in the serum of oil- or CCl₄-treated Tet2^WT and Tet2^ΔMye mice after anti–IL-1β Ab treatment (n = 4–5 for each group). (M and N) Effect of anti–IL-1β Ab treatment on the mRNA levels of Acta2 and Col1a1 (n = 5 for each group). (O) Picrosirius red staining of collagen deposition in the liver tissue of Tet2^WT-CCl₄ and Tet2^ΔMye-CCl₄ mice after treatment of anti–IL-1β Ab (n = 5 for each group). Data are representative of at least two independent experiments with similar results (B, C, and E–O). All data are shown as mean ± SD and were analyzed by two-tailed, paired Student’s t test (B, C, and H, and I) or two-way ANOVA with Sidak’s multiple comparison test (E–G and J–N). ***P < 0.001; **P < 0.01; *P < 0.05; P > 0.05 not significant (ns).