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rabbit anti arg 1  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc rabbit anti arg 1
    Rabbit Anti Arg 1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 728 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/arginase+1/Arginase-1+XP+Rabbit+mAb/pmc12808913-93-40-43
    Average 97 stars, based on 728 article reviews
    rabbit anti arg 1 - by Bioz Stars, 2026-09
    97/100 stars

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    Related Articles

    Incubation:

    Article Title: 4-Octyl Itaconate Promotes Diabetic Wound Healing by Enhancing Pro-Resolving Macrophages via the Efferocytosis-MCT1-Lactate-GPR132 Pathway and Macrophage-Independent Synergistic Effects.
    Article Snippet: .. Tissues were then incubated with primary antibodies against F4/80 (Santa Cruz Biotechnology, Santa Cruz, CA, USA; sc-377009), inducible nitric oxide synthase (iNOS; Cell Signaling Technology, Danvers, MA, USA; 13120), arginase 1 (Arg-1; Cell Signaling Technology; #93668), CD31 (Beyotime Biotechnology, Shanghai, China; AF6408), MCT1 (Proteintech, Rosemont, IL, USA; 20139-1-AP) overnight at 4°C. followed by 1 hour of incubation with fluorescent dye-labeled secondary antibodies (Beyotime Biotechnology; A0428 or A0453). ..

    Article Title: Combined strategy of dual-module cerium nanosystem composite extracellular vesicles regulate ROS in the tissue microenvironment to promote periodontitis recovery
    Article Snippet: A total of 20 μg of protein per sample was separated by SDS-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Germany). .. The membranes were blocked with 5 % bovine serum albumin (BSA, MP Biomedicals, USA) at room temperature for 2 h, then incubated overnight at 4 °C with primary antibodies against iNOS (13120S, Cell Signaling Technology, USA), Arginase-1 (93668S, Cell Signaling Technology, USA), and β-actin (4967S, Cell Signaling Technology, USA). .. After washing with TBST buffer, the membranes were incubated with corresponding secondary antibodies at room temperature for 1 h. Finally, protein bands were visualized using a ChemiDoc chemiluminescence detection system (Bio-Rad, China).

    Article Title: Casbane diterpenoids from Euphorbiaceae suppresse neuroinflammation by regulating microglia polarization via inhibiting NF-κB signaling.
    Article Snippet: Phytochemical investigation of Triadica rotundifolia and Neoshirakia japonica resulted in the discovery of eight new casbane diterpenoid derivatives.. The structures of all isolates were structurally characterized by comprehensive spectroscopic techniques, including UV, IR, NMR, MS, together with X-ray diffraction.. Notably, compound 1 (6S,12R-dihydroxy-1S,2R-casba-3E,7E,10E-trien-5-one) exhibited potent inhibition of NO production in LPS-stimulated microglia.

    Western Blot:

    Article Title: Epimedin C attenuates airway inflammation and remodeling in Asthma by intervening M2 macrophage polarization via modulating the PI3K/Akt/mTOR signaling pathway.
    Article Snippet: .. The following primary and secondary antibodies were used for western blotting: rabbit antibodies from Cell Signaling Technology (CST; MA, USA) against Vimentin (#5741), α -smooth muscle actin (α-SMA; #19245), Arginase-1(Arg-1; #93668), CD206 (#91992), phosphorylated (p) -PI3K (#17366), p-Akt (#4060), and p-mTOR (#5536); mouse antibodies from CST against PI3K (#13666), Akt (#2920), and mTOR (#4517); and a chicken antibody against β-actin (SAB3500350) from Sigma-Aldrich. .. Corresponding horseradish peroxidase (HRP) -conjugated secondary antibodies were goat anti-rabbit IgG (GB23204, Servicebio Biology, Wuhan, China), goat anti-mouse IgG (GB23301, Servicebio), and goat anti-chicken IgY (A16054, Thermo Fisher, MA, USA).

    In Vivo:

    Article Title: Casbane diterpenoids from Euphorbiaceae suppresse neuroinflammation by regulating microglia polarization via inhibiting NF-κB signaling.
    Article Snippet: Phytochemical investigation of Triadica rotundifolia and Neoshirakia japonica resulted in the discovery of eight new casbane diterpenoid derivatives.. The structures of all isolates were structurally characterized by comprehensive spectroscopic techniques, including UV, IR, NMR, MS, together with X-ray diffraction.. Notably, compound 1 (6S,12R-dihydroxy-1S,2R-casba-3E,7E,10E-trien-5-one) exhibited potent inhibition of NO production in LPS-stimulated microglia.

    Blocking Assay:

    Article Title: Casbane diterpenoids from Euphorbiaceae suppresse neuroinflammation by regulating microglia polarization via inhibiting NF-κB signaling.
    Article Snippet: Phytochemical investigation of Triadica rotundifolia and Neoshirakia japonica resulted in the discovery of eight new casbane diterpenoid derivatives.. The structures of all isolates were structurally characterized by comprehensive spectroscopic techniques, including UV, IR, NMR, MS, together with X-ray diffraction.. Notably, compound 1 (6S,12R-dihydroxy-1S,2R-casba-3E,7E,10E-trien-5-one) exhibited potent inhibition of NO production in LPS-stimulated microglia.

    Binding Assay:

    Article Title: Casbane diterpenoids from Euphorbiaceae suppresse neuroinflammation by regulating microglia polarization via inhibiting NF-κB signaling.
    Article Snippet: Phytochemical investigation of Triadica rotundifolia and Neoshirakia japonica resulted in the discovery of eight new casbane diterpenoid derivatives.. The structures of all isolates were structurally characterized by comprehensive spectroscopic techniques, including UV, IR, NMR, MS, together with X-ray diffraction.. Notably, compound 1 (6S,12R-dihydroxy-1S,2R-casba-3E,7E,10E-trien-5-one) exhibited potent inhibition of NO production in LPS-stimulated microglia.

    Staining:

    Article Title: Loss of Early Growth Response Protein 1 in the Liver Leads to Hepatic Lipid Accumulation Driven by an Imbalance Between Fatty Acid β-Oxidation and Oxidative Phosphorylation
    Article Snippet: .. Sections of liver tissue were stained with hematoxylin and eosin (H&E), glutamine synthetase (Abcam, #ab176562), and arginase 1 (Cell Signaling, #93668S) according to standard protocols. .. Frozen liver tissues were cut into 5 μm sections and used for Oil Red O staining (Sigma Aldrich, #00625).



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    ( A to I ) Serpine1 +/+ and Serpine1 −/− mice were orthotopically implanted with pancreatic cancer cells (7940B), and tumors were collected at day 21. [(A) and (B)] Flow cytometry quantification of frequencies of total T cells (CD45 + CD3 + ) (A) and conventional dendritic cells (CD45 + F4/80 − CD11c + MHCII + ) (B) among live cells in orthotopic tumors. (C) Co-IF staining for <t>Arg1</t> (green), F4/80 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 3 FOV per animal). (D) Quantitative RT-PCR analysis of Arg1 in BMDMs treated with vehicle or recombinant PAI1 (rPAI1) for 20 hours. Each dot represents individual primary BMDM lines. (E) Flow cytometry quantification of frequency of CD8 + T cells (CD45 + CD3 + CD8 + ) among live cells in orthotopic tumors. [(F) and (G)] Flow cytometry quantification of frequencies and absolute numbers of GZMB + CD8 + T cells (F) and Ki67 + CD8 + T cells (G) following a 5-hour ex vivo PMA/ionomycin stimulation of tumor digests. (H) Co-IF staining for GZMB (green), CD8 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). (I) Co-IF staining for Ki67 (red), CD8 (green), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). ( J ) Experimental design for CD8 + T cell depletion. Mice received control immunoglobulin G (IgG) or anti-CD8 antibody twice per week, starting 5 days postimplantation (7940B). Tumors were harvested at day 21. ( K ) Tumor weight of orthotopic tumors with CD8⁺ T cell antibody-mediated depletion. Symbols in (A) to (I) and (K) represent individual mice. Data are means ± SEM. P values were determined by two-tailed unpaired t test [(A), (B), right of (C), (D), (E) to (G), and right of (H)], Mann-Whitney test [left of (C), left of (H), and (I)], and Mann-Whitney test with Holm-Sidak post hoc (K).
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    ( A to I ) Serpine1 +/+ and Serpine1 −/− mice were orthotopically implanted with pancreatic cancer cells (7940B), and tumors were collected at day 21. [(A) and (B)] Flow cytometry quantification of frequencies of total T cells (CD45 + CD3 + ) (A) and conventional dendritic cells (CD45 + F4/80 − CD11c + MHCII + ) (B) among live cells in orthotopic tumors. (C) Co-IF staining for <t>Arg1</t> (green), F4/80 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 3 FOV per animal). (D) Quantitative RT-PCR analysis of Arg1 in BMDMs treated with vehicle or recombinant PAI1 (rPAI1) for 20 hours. Each dot represents individual primary BMDM lines. (E) Flow cytometry quantification of frequency of CD8 + T cells (CD45 + CD3 + CD8 + ) among live cells in orthotopic tumors. [(F) and (G)] Flow cytometry quantification of frequencies and absolute numbers of GZMB + CD8 + T cells (F) and Ki67 + CD8 + T cells (G) following a 5-hour ex vivo PMA/ionomycin stimulation of tumor digests. (H) Co-IF staining for GZMB (green), CD8 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). (I) Co-IF staining for Ki67 (red), CD8 (green), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). ( J ) Experimental design for CD8 + T cell depletion. Mice received control immunoglobulin G (IgG) or anti-CD8 antibody twice per week, starting 5 days postimplantation (7940B). Tumors were harvested at day 21. ( K ) Tumor weight of orthotopic tumors with CD8⁺ T cell antibody-mediated depletion. Symbols in (A) to (I) and (K) represent individual mice. Data are means ± SEM. P values were determined by two-tailed unpaired t test [(A), (B), right of (C), (D), (E) to (G), and right of (H)], Mann-Whitney test [left of (C), left of (H), and (I)], and Mann-Whitney test with Holm-Sidak post hoc (K).
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    ( A to I ) Serpine1 +/+ and Serpine1 −/− mice were orthotopically implanted with pancreatic cancer cells (7940B), and tumors were collected at day 21. [(A) and (B)] Flow cytometry quantification of frequencies of total T cells (CD45 + CD3 + ) (A) and conventional dendritic cells (CD45 + F4/80 − CD11c + MHCII + ) (B) among live cells in orthotopic tumors. (C) Co-IF staining for <t>Arg1</t> (green), F4/80 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 3 FOV per animal). (D) Quantitative RT-PCR analysis of Arg1 in BMDMs treated with vehicle or recombinant PAI1 (rPAI1) for 20 hours. Each dot represents individual primary BMDM lines. (E) Flow cytometry quantification of frequency of CD8 + T cells (CD45 + CD3 + CD8 + ) among live cells in orthotopic tumors. [(F) and (G)] Flow cytometry quantification of frequencies and absolute numbers of GZMB + CD8 + T cells (F) and Ki67 + CD8 + T cells (G) following a 5-hour ex vivo PMA/ionomycin stimulation of tumor digests. (H) Co-IF staining for GZMB (green), CD8 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). (I) Co-IF staining for Ki67 (red), CD8 (green), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). ( J ) Experimental design for CD8 + T cell depletion. Mice received control immunoglobulin G (IgG) or anti-CD8 antibody twice per week, starting 5 days postimplantation (7940B). Tumors were harvested at day 21. ( K ) Tumor weight of orthotopic tumors with CD8⁺ T cell antibody-mediated depletion. Symbols in (A) to (I) and (K) represent individual mice. Data are means ± SEM. P values were determined by two-tailed unpaired t test [(A), (B), right of (C), (D), (E) to (G), and right of (H)], Mann-Whitney test [left of (C), left of (H), and (I)], and Mann-Whitney test with Holm-Sidak post hoc (K).
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    ( A to I ) Serpine1 +/+ and Serpine1 −/− mice were orthotopically implanted with pancreatic cancer cells (7940B), and tumors were collected at day 21. [(A) and (B)] Flow cytometry quantification of frequencies of total T cells (CD45 + CD3 + ) (A) and conventional dendritic cells (CD45 + F4/80 − CD11c + MHCII + ) (B) among live cells in orthotopic tumors. (C) Co-IF staining for <t>Arg1</t> (green), F4/80 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 3 FOV per animal). (D) Quantitative RT-PCR analysis of Arg1 in BMDMs treated with vehicle or recombinant PAI1 (rPAI1) for 20 hours. Each dot represents individual primary BMDM lines. (E) Flow cytometry quantification of frequency of CD8 + T cells (CD45 + CD3 + CD8 + ) among live cells in orthotopic tumors. [(F) and (G)] Flow cytometry quantification of frequencies and absolute numbers of GZMB + CD8 + T cells (F) and Ki67 + CD8 + T cells (G) following a 5-hour ex vivo PMA/ionomycin stimulation of tumor digests. (H) Co-IF staining for GZMB (green), CD8 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). (I) Co-IF staining for Ki67 (red), CD8 (green), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). ( J ) Experimental design for CD8 + T cell depletion. Mice received control immunoglobulin G (IgG) or anti-CD8 antibody twice per week, starting 5 days postimplantation (7940B). Tumors were harvested at day 21. ( K ) Tumor weight of orthotopic tumors with CD8⁺ T cell antibody-mediated depletion. Symbols in (A) to (I) and (K) represent individual mice. Data are means ± SEM. P values were determined by two-tailed unpaired t test [(A), (B), right of (C), (D), (E) to (G), and right of (H)], Mann-Whitney test [left of (C), left of (H), and (I)], and Mann-Whitney test with Holm-Sidak post hoc (K).
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    ( A to I ) Serpine1 +/+ and Serpine1 −/− mice were orthotopically implanted with pancreatic cancer cells (7940B), and tumors were collected at day 21. [(A) and (B)] Flow cytometry quantification of frequencies of total T cells (CD45 + CD3 + ) (A) and conventional dendritic cells (CD45 + F4/80 − CD11c + MHCII + ) (B) among live cells in orthotopic tumors. (C) Co-IF staining for <t>Arg1</t> (green), F4/80 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 3 FOV per animal). (D) Quantitative RT-PCR analysis of Arg1 in BMDMs treated with vehicle or recombinant PAI1 (rPAI1) for 20 hours. Each dot represents individual primary BMDM lines. (E) Flow cytometry quantification of frequency of CD8 + T cells (CD45 + CD3 + CD8 + ) among live cells in orthotopic tumors. [(F) and (G)] Flow cytometry quantification of frequencies and absolute numbers of GZMB + CD8 + T cells (F) and Ki67 + CD8 + T cells (G) following a 5-hour ex vivo PMA/ionomycin stimulation of tumor digests. (H) Co-IF staining for GZMB (green), CD8 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). (I) Co-IF staining for Ki67 (red), CD8 (green), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). ( J ) Experimental design for CD8 + T cell depletion. Mice received control immunoglobulin G (IgG) or anti-CD8 antibody twice per week, starting 5 days postimplantation (7940B). Tumors were harvested at day 21. ( K ) Tumor weight of orthotopic tumors with CD8⁺ T cell antibody-mediated depletion. Symbols in (A) to (I) and (K) represent individual mice. Data are means ± SEM. P values were determined by two-tailed unpaired t test [(A), (B), right of (C), (D), (E) to (G), and right of (H)], Mann-Whitney test [left of (C), left of (H), and (I)], and Mann-Whitney test with Holm-Sidak post hoc (K).
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    Image Search Results


    ( A to I ) Serpine1 +/+ and Serpine1 −/− mice were orthotopically implanted with pancreatic cancer cells (7940B), and tumors were collected at day 21. [(A) and (B)] Flow cytometry quantification of frequencies of total T cells (CD45 + CD3 + ) (A) and conventional dendritic cells (CD45 + F4/80 − CD11c + MHCII + ) (B) among live cells in orthotopic tumors. (C) Co-IF staining for Arg1 (green), F4/80 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 3 FOV per animal). (D) Quantitative RT-PCR analysis of Arg1 in BMDMs treated with vehicle or recombinant PAI1 (rPAI1) for 20 hours. Each dot represents individual primary BMDM lines. (E) Flow cytometry quantification of frequency of CD8 + T cells (CD45 + CD3 + CD8 + ) among live cells in orthotopic tumors. [(F) and (G)] Flow cytometry quantification of frequencies and absolute numbers of GZMB + CD8 + T cells (F) and Ki67 + CD8 + T cells (G) following a 5-hour ex vivo PMA/ionomycin stimulation of tumor digests. (H) Co-IF staining for GZMB (green), CD8 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). (I) Co-IF staining for Ki67 (red), CD8 (green), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). ( J ) Experimental design for CD8 + T cell depletion. Mice received control immunoglobulin G (IgG) or anti-CD8 antibody twice per week, starting 5 days postimplantation (7940B). Tumors were harvested at day 21. ( K ) Tumor weight of orthotopic tumors with CD8⁺ T cell antibody-mediated depletion. Symbols in (A) to (I) and (K) represent individual mice. Data are means ± SEM. P values were determined by two-tailed unpaired t test [(A), (B), right of (C), (D), (E) to (G), and right of (H)], Mann-Whitney test [left of (C), left of (H), and (I)], and Mann-Whitney test with Holm-Sidak post hoc (K).

    Journal: Science Advances

    Article Title: A stromal PAI1-tPA axis orchestrates immunosuppression in pancreatic cancer

    doi: 10.1126/sciadv.aea6734

    Figure Lengend Snippet: ( A to I ) Serpine1 +/+ and Serpine1 −/− mice were orthotopically implanted with pancreatic cancer cells (7940B), and tumors were collected at day 21. [(A) and (B)] Flow cytometry quantification of frequencies of total T cells (CD45 + CD3 + ) (A) and conventional dendritic cells (CD45 + F4/80 − CD11c + MHCII + ) (B) among live cells in orthotopic tumors. (C) Co-IF staining for Arg1 (green), F4/80 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 3 FOV per animal). (D) Quantitative RT-PCR analysis of Arg1 in BMDMs treated with vehicle or recombinant PAI1 (rPAI1) for 20 hours. Each dot represents individual primary BMDM lines. (E) Flow cytometry quantification of frequency of CD8 + T cells (CD45 + CD3 + CD8 + ) among live cells in orthotopic tumors. [(F) and (G)] Flow cytometry quantification of frequencies and absolute numbers of GZMB + CD8 + T cells (F) and Ki67 + CD8 + T cells (G) following a 5-hour ex vivo PMA/ionomycin stimulation of tumor digests. (H) Co-IF staining for GZMB (green), CD8 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). (I) Co-IF staining for Ki67 (red), CD8 (green), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). ( J ) Experimental design for CD8 + T cell depletion. Mice received control immunoglobulin G (IgG) or anti-CD8 antibody twice per week, starting 5 days postimplantation (7940B). Tumors were harvested at day 21. ( K ) Tumor weight of orthotopic tumors with CD8⁺ T cell antibody-mediated depletion. Symbols in (A) to (I) and (K) represent individual mice. Data are means ± SEM. P values were determined by two-tailed unpaired t test [(A), (B), right of (C), (D), (E) to (G), and right of (H)], Mann-Whitney test [left of (C), left of (H), and (I)], and Mann-Whitney test with Holm-Sidak post hoc (K).

    Article Snippet: The following antibodies were used: Arg1 (D4E3M, 1:400, no. 93668S, Cell Signaling), cleaved caspase-3 (polyclonal, 1:100, no. 9661S, Cell Signaling), CD8 (D4W2Z, 1:200, no. 98941S, Cell Signaling), ECAD (36, 1:1000, no. 610181, BD Biosciences), F4/80 (D2S9R, 1:200, no. 70076S, Cell Signaling), GZMB (D6E9W, 1:100, no. 46890S, Cell Signaling), Ki67 (SP6, 1:200, no. MA5-14520, Invitrogen), PAI1 (polyclonal, 1:1000, no. IRBAHUPAI1AP100UG, Innovative Research), PanCK (monoclonal antibody cocktail, 1:400, no. NBP2-44368-0.1 mg, Novus), PDGFRα/β (Y92, 1:100, no. AB32570, Abcam), and tPA (polyclonal, 1:250, no. 10147-1-AP, Proteintech).

    Techniques: Flow Cytometry, Staining, Quantitative RT-PCR, Recombinant, Ex Vivo, Control, Two Tailed Test, MANN-WHITNEY

    ( A ) Experimental outline of murine PDAC model. Plat +/+ and Plat −/− mice were orthotopically implanted with pancreatic cancer cells (7940B), and tumors were collected 21 days postimplantation. ( B ) Tumor weight of orthotopic tumors harvested from Plat +/+ and Plat −/− mice. ( C ) ELISA of tPA in tissue supernatants from pancreata of Plat +/+ mice (no tumor) and orthotopic tumors from Plat +/+ and Plat −/− mice. ( D ) Picrosirius red staining of orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). ( E ) Co-IF staining for cCasp3 (red), ECAD (green), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 4 FOV per animal). ( F to H ) Flow cytometry quantification of frequencies of total T cells (F), CD8 + T cells (G), and conventional dendritic cells (H) among live cells in orthotopic tumors. ( I ) Co-IF staining for Arg1 (green), F4/80 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 3 FOV per animal). Symbols in (B) to (I) represent individual mice. Data are means ± SEM. P values were determined by Mann-Whitney test [(B) and (F)], two-way ANOVA with Holm-Sidak post hoc (C), and two-tailed unpaired t test [(D), (E), and (G) to (I)].

    Journal: Science Advances

    Article Title: A stromal PAI1-tPA axis orchestrates immunosuppression in pancreatic cancer

    doi: 10.1126/sciadv.aea6734

    Figure Lengend Snippet: ( A ) Experimental outline of murine PDAC model. Plat +/+ and Plat −/− mice were orthotopically implanted with pancreatic cancer cells (7940B), and tumors were collected 21 days postimplantation. ( B ) Tumor weight of orthotopic tumors harvested from Plat +/+ and Plat −/− mice. ( C ) ELISA of tPA in tissue supernatants from pancreata of Plat +/+ mice (no tumor) and orthotopic tumors from Plat +/+ and Plat −/− mice. ( D ) Picrosirius red staining of orthotopic tumors, and corresponding quantification ( n = 5 FOV per animal). ( E ) Co-IF staining for cCasp3 (red), ECAD (green), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 4 FOV per animal). ( F to H ) Flow cytometry quantification of frequencies of total T cells (F), CD8 + T cells (G), and conventional dendritic cells (H) among live cells in orthotopic tumors. ( I ) Co-IF staining for Arg1 (green), F4/80 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 3 FOV per animal). Symbols in (B) to (I) represent individual mice. Data are means ± SEM. P values were determined by Mann-Whitney test [(B) and (F)], two-way ANOVA with Holm-Sidak post hoc (C), and two-tailed unpaired t test [(D), (E), and (G) to (I)].

    Article Snippet: The following antibodies were used: Arg1 (D4E3M, 1:400, no. 93668S, Cell Signaling), cleaved caspase-3 (polyclonal, 1:100, no. 9661S, Cell Signaling), CD8 (D4W2Z, 1:200, no. 98941S, Cell Signaling), ECAD (36, 1:1000, no. 610181, BD Biosciences), F4/80 (D2S9R, 1:200, no. 70076S, Cell Signaling), GZMB (D6E9W, 1:100, no. 46890S, Cell Signaling), Ki67 (SP6, 1:200, no. MA5-14520, Invitrogen), PAI1 (polyclonal, 1:1000, no. IRBAHUPAI1AP100UG, Innovative Research), PanCK (monoclonal antibody cocktail, 1:400, no. NBP2-44368-0.1 mg, Novus), PDGFRα/β (Y92, 1:100, no. AB32570, Abcam), and tPA (polyclonal, 1:250, no. 10147-1-AP, Proteintech).

    Techniques: Enzyme-linked Immunosorbent Assay, Staining, Flow Cytometry, MANN-WHITNEY, Two Tailed Test

    ( A ) Experimental outline of murine PDAC model. WT, Serpine1 −/− , and Serpine1 −/− ; Plat −/− mice were orthotopically implanted with pancreatic cancer cells (7940B), and tumors were collected 21 days postimplantation. ( B ) Tumor weight of orthotopic tumors harvested from WT, Serpine1 −/− , and Serpine1 −/− ; Plat −/− mice. ( C ) IHC staining for CD8 in orthotopic tumors, and corresponding quantification ( n = 3 FOV per animal). ( D ) Co-IF staining for Arg1 (green), F4/80 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 3 FOV per animal). Symbols in (B) to (D) represent individual mice. Data are means ± SEM. P values were determined by one-way ANOVA with Holm-Sidak post hoc [(B) to (D)]. ( E ) Working model. Left: Hypoxia stabilizes HIF proteins, inducing PAI1 expression in pancreatic CAFs. Although tPA levels are elevated in PDAC, PAI1 predominantly inhibits its activity, thereby suppressing antitumor immunity. Middle: Elimination of stromal PAI1 restores tPA activity, enhancing antitumor CD8 + T cell responses, accompanied by alleviation of immunosuppressive TAM phenotypes and increased dendritic cell (DC) infiltration. Stromal PAI1-driven tumor growth depends on CD8 + T cells. Right: Removal of stromal tPA abolishes residual tPA activity, further promoting an immunosuppressive TME and tumor growth.

    Journal: Science Advances

    Article Title: A stromal PAI1-tPA axis orchestrates immunosuppression in pancreatic cancer

    doi: 10.1126/sciadv.aea6734

    Figure Lengend Snippet: ( A ) Experimental outline of murine PDAC model. WT, Serpine1 −/− , and Serpine1 −/− ; Plat −/− mice were orthotopically implanted with pancreatic cancer cells (7940B), and tumors were collected 21 days postimplantation. ( B ) Tumor weight of orthotopic tumors harvested from WT, Serpine1 −/− , and Serpine1 −/− ; Plat −/− mice. ( C ) IHC staining for CD8 in orthotopic tumors, and corresponding quantification ( n = 3 FOV per animal). ( D ) Co-IF staining for Arg1 (green), F4/80 (red), and DAPI (blue) in orthotopic tumors, and corresponding quantification ( n = 3 FOV per animal). Symbols in (B) to (D) represent individual mice. Data are means ± SEM. P values were determined by one-way ANOVA with Holm-Sidak post hoc [(B) to (D)]. ( E ) Working model. Left: Hypoxia stabilizes HIF proteins, inducing PAI1 expression in pancreatic CAFs. Although tPA levels are elevated in PDAC, PAI1 predominantly inhibits its activity, thereby suppressing antitumor immunity. Middle: Elimination of stromal PAI1 restores tPA activity, enhancing antitumor CD8 + T cell responses, accompanied by alleviation of immunosuppressive TAM phenotypes and increased dendritic cell (DC) infiltration. Stromal PAI1-driven tumor growth depends on CD8 + T cells. Right: Removal of stromal tPA abolishes residual tPA activity, further promoting an immunosuppressive TME and tumor growth.

    Article Snippet: The following antibodies were used: Arg1 (D4E3M, 1:400, no. 93668S, Cell Signaling), cleaved caspase-3 (polyclonal, 1:100, no. 9661S, Cell Signaling), CD8 (D4W2Z, 1:200, no. 98941S, Cell Signaling), ECAD (36, 1:1000, no. 610181, BD Biosciences), F4/80 (D2S9R, 1:200, no. 70076S, Cell Signaling), GZMB (D6E9W, 1:100, no. 46890S, Cell Signaling), Ki67 (SP6, 1:200, no. MA5-14520, Invitrogen), PAI1 (polyclonal, 1:1000, no. IRBAHUPAI1AP100UG, Innovative Research), PanCK (monoclonal antibody cocktail, 1:400, no. NBP2-44368-0.1 mg, Novus), PDGFRα/β (Y92, 1:100, no. AB32570, Abcam), and tPA (polyclonal, 1:250, no. 10147-1-AP, Proteintech).

    Techniques: Immunohistochemistry, Staining, Expressing, Activity Assay