Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Epigenetic metabolite lipid nanoparticles alleviate venous thrombosis via bone marrow reprogramming

doi: 10.1016/j.bvth.2025.100086

Figure Lengend Snippet: Immunomodulatory effects of ITA-LNP in vitro and in vivo. (A) Immunoblot probing for IL-1β in BMDMs treated with 100 ng/mL LPS in the presence of various doses of ITA-LNPs. (B) Schematic of the experiments in C57BL/6 mice subjected to LPS-induced endotoxemia. (C) Kaplan-Meier survival analysis after LPS-induced (15 mg/kg) endotoxemia in C57BL/6 mice pretreated with Ctrl/ITA-LNPs or anti–IL-1β mAb. (D) Rectal temperature measured in the mice from panel C before and 6 hours after LPS injections. (E-F) The levels of IL-1β and IL-6 in the serum from mice of a separate cohort injected as in panel B but with a sublethal dose of 8 mg/kg LPS; n = 10 to 15 mice per group. (G) NETs in primary human neutrophils incubated for 5 hours in vitro with 50 μM LNPs as indicated and in the presence of 100 nM PMA followed by immunofluorescence analysis; n = 6 for Ctrl-LNP and ITA-LNP without PMA; n = 8 for Ctrl-LNP with PMA; n = 9 for ITA-LNP with PMA; and n = 4 for PBS groups with or without PMA. (H) Cytokine analysis in supernatants from experiments in panel G. Statistical analysis is through pairwise t test with Holm post hoc test . N.d., not detected; PBS, phosphate-buffered saline; ROI, region of interest; T, temperature.

Article Snippet: C57BL/6 male mice (10 weeks old, n = 15 per group) were IV injected with 50 mg/kg ITA- or control (Ctrl)-LNPs or 50 mg/kg anti-mouse IL-1β monoclonal antibody (clone 7E3, InvivoGen mil1b-mab9-10).

Techniques: In Vitro, In Vivo, Western Blot, Injection, Incubation, Immunofluorescence, Saline

Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Epigenetic metabolite lipid nanoparticles alleviate venous thrombosis via bone marrow reprogramming

doi: 10.1016/j.bvth.2025.100086

Figure Lengend Snippet: ITA-LNPs alleviate venous thrombosis in IVC ligation model. (A) Schematic of the experiments. WT mice IV pretreated (50 mg/kg, bolus) with ITA- or Ctrl-LNPs were subjected to IVC ligation surgery followed by the isolation of the blood clots; n = 6 for Ctrl-LNP and n = 8 for ITA-LNP. (B) Gross pathology with clot weight quantification. (C) Hematoxylin and eosin staining of blood clot sections from these experiments. (D) Immunofluorescence staining of the same sections using Ly6G/C mAb (Red) and nuclei (blue). (E) MPO (top, alkaline phosphatase immunohistochemistry) and IL-1β (bottom, immunofluorescence) staining in the same clots. (F) Quantification of the staining in panel E. For each biological replicate (n = 6 for Ctrl-LNP and n = 8 for ITA = LNP), 3 to 4 histopathological locations at different depths (100 μm apart) were analyzed. (G) NET staining in the same sections using triple-positive identification through colocalization of the signals from MPO (green), H4Cit3 (red), and DNA (DAPI; blue). (H) Quantification of the staining in panel G. Quantification was accomplished using 2 to 3 different histopathological locations and the same aforementioned biological replicates. Statistical analysis is through a 2-tailed t test. DAPI, 4′,6-diamidino-2-phenylindole; H4Cit3, citrullinated histone H4; MPO, myeloperoxidase; WT, wild type.

Article Snippet: C57BL/6 male mice (10 weeks old, n = 15 per group) were IV injected with 50 mg/kg ITA- or control (Ctrl)-LNPs or 50 mg/kg anti-mouse IL-1β monoclonal antibody (clone 7E3, InvivoGen mil1b-mab9-10).

Techniques: Ligation, Isolation, Staining, Immunofluorescence, Immunohistochemistry

Journal: Nature Communications

Article Title: Type 1 innate lymphoid cell–immature neutrophil axis suppresses acute tissue inflammation

doi: 10.1038/s41467-025-61504-8

Figure Lengend Snippet: a Plasma concentrations of IL-10 in WT and Ifng –/– mice before (naive) ( n = 9 and 4, respectively) and at 6 h after ( n = 5 and 5, respectively) liver IR. b Immunoblotting analysis of IL-10 and GAPDH (left) and relative expressions of IL-10 normalized to GAPDH (right) in the liver of WT and Ifng –/– mice before (naive) ( n = 3 and 4, respectively) and at 6 h after ( n = 4 and 4, respectively) liver IR. c –e Plasma concentrations of ALT ( c ) and necrotic ( d ) and hemorrhage ( e ) areas in the liver of WT and Ifng –/– mice treated with isotype control Ig or anti-IL-10 mAb before and at 6 h after liver IR ( n = 3 in naive WT mice and n = 4 in the other groups). f Relative quantity (RQ) of Il10 mRNA expression, normalized to Actb , in inflammatory monocytes (iMo), neutrophils, conventional dendritic cells (cDC), and Kupper cells in the liver of WT and Ifng –/– mice before (naive) and 5 h after liver IR ( n = 7 in each group). n.d.: not detected. g Plasma IL-10 concentration in WT and Ifng –/– mice injected with or without anti-Ly6G mAb or control mAb (cIg) 24 h before IR ( n = 6). h Il10 mRNA expression, normalized to Actb , in CD49d + and CD49d – neutrophils in the liver and blood circulation of WT mice ( n = 4 in each group of neutrophils in naive mice, n = 5 in CD49d + neutrophils in the liver after IRI, and n = 6 in the other groups). i Il10 mRNA expression, normalized to Actb , in CD49d – imNeu after stimulation with LPS or supernatants of liver homogenates before (naive liver) and at 3 h after IRI (IRI liver) ( n = 6). A selective TLR4 antagonist TAK-242 (TAK) or vehicle (Veh) was pretreated for 1 h before stimulation with liver homogenates. C57BL/6J mice were used ( a – i ). Data are pooled from two independent experiments and represented as the mean ± SD ( a – i ). Statistical analysis was performed using one-way ANOVA, Tukey’s HSD test.

Article Snippet: To block cytokines signaling, 100 μg of neutralizing mAbs against mouse IFN-γ (clone R4-6A2, Bio X Cell, New Hampshire, USA) or mouse IL-10 (clone JES5-2A5, Bio X Cell, New Hampshire, USA) were injected into mice intraperitoneally before reperfusion.

Techniques: Clinical Proteomics, Western Blot, Control, Expressing, Concentration Assay, Injection

Journal: Nature Communications

Article Title: Type 1 innate lymphoid cell–immature neutrophil axis suppresses acute tissue inflammation

doi: 10.1038/s41467-025-61504-8

Figure Lengend Snippet: a –d Body weight change ( a , c ) and mortality ( b , d ) in WT, Ifng –/– , or Ahnak –/– mice that underwent or did not cecal ligation and puncture (CLP) on day 0. e Body weight change in Ifng –/– mice that received IFN-γ ( n = 6) or vehicle ( n = 6) injection one day before CLP on day 0. f Body weight change in naive Rag2 –/– Il2rg –/– mice ( n = 4) or those that had been i.p. transferred or not with WT ( n = 7 and 4, respectively) or Ifng –/– ( n = 7) ILC1 one day before CLP on day 0. g The numbers of CD49d – imNeu and CD49d + mNeu in the blood circulation at 6 days after CLP in WT and Ifng –/– mice ( n = 5 in each group). h Il10 mRNA expression in CD49d – imNeu and CD49d + mNeu in the blood circulation and peritoneal cavity in naive mice ( n = 9) or mice at 7 days after CLP ( n = 8). i Mortality in mice treated with control mAb (cIg) or anti-IL-10 mAb one day before CLP on day 0 ( n = 9 in each group). j Body weight change in mice treated with control mAb ( n = 9), anti-IFN-γ mAb ( n = 9), anti-IL-10 mAb ( n = 9), or the cocktail of anti-IFN-γ mAb and anti-IL-10 mAb ( n = 10) one day before CLP on day 0. C57BL/6J female mice were used ( a – j ). Data are pooled from three ( a , b , h , i and j ) and two ( c – g ) independent experiments and represented as the mean ± SD. Statistical analyses were performed using one-way ANOVA, Tukey’s HSD test. P values, WT-CLP vs Ifng –/– -CLP ( a ), WT-CLP vs Ahnak –/– -CLP ( c ), Ifng –/– -vehcle vs Ifng –/– -IFN-γ ( e ), and WT-ILC1 vs Ifng –/– -ILC1 ( f ).

Article Snippet: To block cytokines signaling, 100 μg of neutralizing mAbs against mouse IFN-γ (clone R4-6A2, Bio X Cell, New Hampshire, USA) or mouse IL-10 (clone JES5-2A5, Bio X Cell, New Hampshire, USA) were injected into mice intraperitoneally before reperfusion.

Techniques: Ligation, Injection, Expressing, Control