anti il 1β  (InvivoGen)

Journal: Nature

Article Title: Bidirectional CRISPR screens decode a GLIS3-dependent fibrotic cell circuit

doi: 10.1038/s41586-025-09907-x

Figure Lengend Snippet: (a) Secreted IL-11 after co-culture of fibroblast knockouts for TGFB1 - and IL1B -related ligands or receptors with TLR2/6-activated macrophages (top) or knockouts in activated macrophage knockouts with fibroblasts (bottom). n=3 cell lines per condition. (b) Z-score heatmap of IAF genes in TGFBR1/2 , IL1R1 CRISPRko fibroblasts co-cultured with TLR2/6-activated macrophages, normalized to HPRT. n=3 cell lines per condition. (c) qPCR of gene knockouts normalized to HPRT . Macrophages: IL-4/IL-13 (10 ng/mL); FSL-1 (10 ng/mL) + ATP (5 mM). Fibroblasts: media. n=2 cell lines per condition. (d) Secreted TGF-β and IL-1β after co-culture of primary human macrophages and fibroblasts (24 hours) ( Methods ). n=3 cell lines per condition. (e) Secreted IL-11 from primary colonic fibroblasts stimulated with TGF-β and/or IL-1β (10 ng/mL, 24 hours). Dashed lines: additive or synergistic response ( Methods ). One-way ANOVA with Tukey’s multiple-comparisons test. n=3 cell lines per condition. (f) Left: immunofluorescence of Il11 mNG colons after intraperitoneal injection with IgG control or dual anti-TGF-β and anti-IL-1β antibodies (100 μL in PBS, 100 μg/mouse). Right: IL-11 mNG cell percentage in all DAPI-imaged cells from two pooled independent experiments. Mice (co-housed, 13-20 weeks) were treated with chronic DSS (2.0%, 35 days). n=7 mice per condition. (g) qPCR quantification of Il11 from lysates from (f) normalized to Eef2 . Kruskal-Wallis test with Dunn’s multiple-comparison test. (h) Total colonic collagen percentage quantification from (f). (i) Quantification of colonic hydroxyproline normalized to total protein from lysates from (f). (j) Percent starting weight of mice from (f). Filled lines represent s.e.m. Linear mixed-effects analysis with Dunnett’s multiple comparison test. (k) Histopathological scoring ( Methods ) of H&E-stained tissues from (f). (l) Colon length measurements from (f). Unless otherwise stated, statistics are by a one-way ANOVA with Dunnet’s multiple comparison test on distinct biological replicates and error bars are the mean ± s.e.m. ns, not significant.

Article Snippet: Il11 mNG mice were administered DSS following the chronic DSS model. At the start of each cycle of DSS administration, mice were IP injected with monoclonal antibodies (mAbs) directed against IgG (BioXCell, #BP0083), anti-TGF-β (BioXCell, #BP0057), anti-IL-1β (Invivogen, #mil1b-mab9-1T), or the combination of anti-TGF-β and anti-IL-1β.

Techniques: Co-Culture Assay, Cell Culture, Immunofluorescence, Injection, Control, Comparison, Staining

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: Biomaterials Research

Article Title: M2 Macrophage-Derived Small Extracellular Vesicles Ameliorate Pyroptosis and Intervertebral Disc Degeneration

doi: 10.34133/bmr.0047

Figure Lengend Snippet: Schematic diagram for M2 macrophage-derived sEVs release from dECM-hydrogels and the subsequent delivery of miR-221-3p to inhibit nucleus pulposus cell pyroptosis via modulation of the PTEN/NLRP3 signaling pathway.

Article Snippet: The primary antibodies and dilutions were as follows: anti-Caspase-1(p20) (Proteintech, China, 22915-1, 1:1000), anti-NLRP3 (Proteintech, China, 19771-1, 1:1000), anti-mIL-1β (Abcam, UK, ab283818, 1:1,000), anti-MMP13 (Proteintech, China, 18165-1, 1:1,000), anti-Aggrecan (Abcam, UK, ab36861, 1:1,000), anti-Collagen II (Abcam, UK, ab188570, 1:1,000), and anti-PTEN (CST, USA, 9559S, 1:1,000).

Techniques: Derivative Assay

Journal: Biomaterials Research

Article Title: M2 Macrophage-Derived Small Extracellular Vesicles Ameliorate Pyroptosis and Intervertebral Disc Degeneration

doi: 10.34133/bmr.0047

Figure Lengend Snippet: The expression of Caspase-1 increases with the degree of IDD. (A) Typical MRI images of II to V Pfirrmann grading. (B) Representative photographs of IHC for Caspase-1-positive cells in degenerated NP tissue (scale bar: 50 μm). (C) Corresponding quantification of positive protein expression levels. Data are presented as the mean ± SD for each group (* P < 0.05).

Article Snippet: The primary antibodies and dilutions were as follows: anti-Caspase-1(p20) (Proteintech, China, 22915-1, 1:1000), anti-NLRP3 (Proteintech, China, 19771-1, 1:1000), anti-mIL-1β (Abcam, UK, ab283818, 1:1,000), anti-MMP13 (Proteintech, China, 18165-1, 1:1,000), anti-Aggrecan (Abcam, UK, ab36861, 1:1,000), anti-Collagen II (Abcam, UK, ab188570, 1:1,000), and anti-PTEN (CST, USA, 9559S, 1:1,000).

Techniques: Expressing

Journal: Biomaterials Research

Article Title: M2 Macrophage-Derived Small Extracellular Vesicles Ameliorate Pyroptosis and Intervertebral Disc Degeneration

doi: 10.34133/bmr.0047

Figure Lengend Snippet: M2-sEVs inhibits H 2 O 2 -induced pyroptosis of NP cells. (A and B) The TUNEL staining and quantitative analysis was used to determine the pyroptosis of NPCs treated with H 2 O 2 (200 μM) with or without M2-sEVs (scale bar: 100 μm). (C) Hoechst33342/PI double fluorescence staining was used to detect the changes of cell membrane permeability after NPCs was treated with H 2 O 2 with or without M2-sEVs (scale bar: 100 μm). (D) Annexin V/PI flow cytometry staining was used to detect PI-positive cells in NPC treated with H 2 O 2 with or without M2-sEVs and quantitative analysis was shown in (E). (F) DCFH-DA staining was used to detect the changes of oxygen content after NPCs was treated with H 2 O 2 with or without M2-sEVs (scale bar: 100 μm). (G) Quantitative analysis of ROS-positive NPCs. The secretion of (H) IL-18 and (I) IL-1β from NPCs treated with H 2 O 2 with or without M2-sEVs, as detected by ELISA. The relative expression levels of Col II, Agg, NLRP3, MMP13, Caspase-1 (p20), and mIL-1β were assessed by (J) RT-qPCR and (K) Western blotting in HNPC treated with H 2 O 2 with or without M2-sEVs. (L) Quantitative analysis of Western blot protein bands. Data are presented as the mean ± SD for each group (* P < 0.05, ** P < 0.01).

Article Snippet: The primary antibodies and dilutions were as follows: anti-Caspase-1(p20) (Proteintech, China, 22915-1, 1:1000), anti-NLRP3 (Proteintech, China, 19771-1, 1:1000), anti-mIL-1β (Abcam, UK, ab283818, 1:1,000), anti-MMP13 (Proteintech, China, 18165-1, 1:1,000), anti-Aggrecan (Abcam, UK, ab36861, 1:1,000), anti-Collagen II (Abcam, UK, ab188570, 1:1,000), and anti-PTEN (CST, USA, 9559S, 1:1,000).

Techniques: TUNEL Assay, Staining, Fluorescence, Membrane, Permeability, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: Biomaterials Research

Article Title: M2 Macrophage-Derived Small Extracellular Vesicles Ameliorate Pyroptosis and Intervertebral Disc Degeneration

doi: 10.34133/bmr.0047

Figure Lengend Snippet: The characterization, biocompatibility, and releases of ECM hydrogel. (A) Photographs of dECM-hydrogels and their ability to form gels at different concentrations. (B) Rheological test results of dECM-hydrogels of different concentrations. (C) Live/Dead staining of NPCs seeded on 1% and 2% dECM-hydrogels. Scale bar: 100 μm. (D) Phalloidin staining of NPCs seeded on 1% and 2% hydrogel. Scale bar: 100 μm. (E) The sustained-release curves of M2-sEVs from 1% dECM-hydrogels. (F) CCK-8 assay results showed that the cell viability of H 2 O 2 -treated cells was rescued by M2-sEVs release from dECM-hydrogels. Data are shown as the mean ± SD for each group (* P < 0.05).

Article Snippet: The primary antibodies and dilutions were as follows: anti-Caspase-1(p20) (Proteintech, China, 22915-1, 1:1000), anti-NLRP3 (Proteintech, China, 19771-1, 1:1000), anti-mIL-1β (Abcam, UK, ab283818, 1:1,000), anti-MMP13 (Proteintech, China, 18165-1, 1:1,000), anti-Aggrecan (Abcam, UK, ab36861, 1:1,000), anti-Collagen II (Abcam, UK, ab188570, 1:1,000), and anti-PTEN (CST, USA, 9559S, 1:1,000).

Techniques: Staining, CCK-8 Assay

Journal: Biomaterials Research

Article Title: M2 Macrophage-Derived Small Extracellular Vesicles Ameliorate Pyroptosis and Intervertebral Disc Degeneration

doi: 10.34133/bmr.0047

Figure Lengend Snippet: Radiography and histological evaluation of ECM hydrogel released M2-sEVs in the rat model. (A) X-ray images of the IVD at 0, 4, and 8 weeks. (B) Representative MRI images of rat coccygeal in the different groups. (C) DHI% of IVD differences among the groups at 4 and 8 weeks. (D) The Pfirrmann grading of the MRI images. (E) Images of H&E, Safranin-O-Fast Green, and Alcian Blue staining in each group (scale bar: 1 mm) and (F) histological scoring shown alongside. (G) Western blots and corresponding quantification of (H) Col II, (I) NLRP3, and (J) Caspase-1(p20) total protein expression in each group. Data are shown as the mean ± SD for each group (* P < 0.05, ** P < 0.01, ***P < 0.001, **** P < 0.0001).

Article Snippet: The primary antibodies and dilutions were as follows: anti-Caspase-1(p20) (Proteintech, China, 22915-1, 1:1000), anti-NLRP3 (Proteintech, China, 19771-1, 1:1000), anti-mIL-1β (Abcam, UK, ab283818, 1:1,000), anti-MMP13 (Proteintech, China, 18165-1, 1:1,000), anti-Aggrecan (Abcam, UK, ab36861, 1:1,000), anti-Collagen II (Abcam, UK, ab188570, 1:1,000), and anti-PTEN (CST, USA, 9559S, 1:1,000).

Techniques: Staining, Western Blot, Expressing