hmp unc2025  (MedChemExpress)

Journal: Science Advances

Article Title: NLRP3 inflammasome constrains liver regeneration through impairing MerTK-mediated macrophage efferocytosis

doi: 10.1126/sciadv.adq5786

Figure Lengend Snippet: ( A ) ELISA analysis of serum IL-6 level ( n = 3 per group). ( B ) ELISA analysis of serum TNF-α level ( n = 3 per group). ( C and D ) Representative flow cytometry analysis plots and the quantification of Ly6C hi or Ly6C lo hepatic macrophages 36 hours after PHx ( n = 3 per group). ( E ) mRNA expression of Ly6C hi or Ly6C lo macrophage markers in isolated hepatic macrophages ( n = 3 per group). iNOS, inducible nitric oxide synthase; Arg-1, arginase-1. ( F ) RNA sequencing (RNA-seq) analysis of isolated hepatic macrophages 36 hours after PHx. Heatmap showed the proinflammatory (M1), reparative (M2), and efferocytosis gene expression profiles in Nlrp3 Δ mye mice and Nlrp3 fl/fl mice. ( G ) Bubble chart showing the top 20 of Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment of the significant up-regulated genes in Nlrp3 Δ mye mice. PPAR, peroxisome proliferator–activated receptor. ( H and I ) mRNA expression of efferocytosis genes in isolated hepatic macrophages 36 hours after PHx ( n = 3 per group). ( J ) Representative fluorescent images and quantification of engulfing ACs (TUNEL) by hepatic macrophages (CD68) in situ from Nlrp3 Δ mye mice and Nlrp3 fl/fl mice 36 hours after PHx (scale bar, 50 μm) ( n = 5 per group). ( K ) Flow cytometry analysis of MerTK expression in isolated hepatic macrophages 36 hours after PHx ( n = 3 per group). ( L ) Representative fluorescent images and quantification of engulfing ACs (pHrodo labeled, green) by BMDMs (F-actin labeled, red) isolated from Nlrp3 Δ mye mice and Nlrp3 fl/fl mice (scale bar, 50 μm) ( n = 5 per group). ( M ) mRNA expression of MerTK and HGF in isolated hepatic macrophages after cocultured with ACs ( n = 3 per group). Data are shown as means ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001. h, hours.

Article Snippet: For MerTK inhibition, UNC2025 (TargetMol, no. T7007) was administrated intraperitoneally to a concentration of 50 mg/kg 2 hours before and 24 hours after PHx.

Techniques: Enzyme-linked Immunosorbent Assay, Flow Cytometry, Expressing, Isolation, RNA Sequencing Assay, TUNEL Assay, In Situ, Labeling

Journal: Science Advances

Article Title: NLRP3 inflammasome constrains liver regeneration through impairing MerTK-mediated macrophage efferocytosis

doi: 10.1126/sciadv.adq5786

Figure Lengend Snippet: The NLRP3 inflammasome exhibited robust activation in macrophages during the initial phase of liver regeneration following 70% PHx. Blocking macrophage NLRP3 significantly enhanced liver regeneration, whereas overexpression of NLRP3 impaired it after PHx. Deficiency in Nlrp3 promoted MerTK-mediated efferocytosis, thereby inducing a pro-reparative Ly6C lo phenotype in macrophages. This image was drawn by the authors.

Article Snippet: For MerTK inhibition, UNC2025 (TargetMol, no. T7007) was administrated intraperitoneally to a concentration of 50 mg/kg 2 hours before and 24 hours after PHx.

Techniques: Activation Assay, Blocking Assay, Over Expression