rat anti mouse il 38 antibody  (R&D Systems)

Journal: Mediators of Inflammation

Article Title: IL-38 Alleviates Inflammation in Sepsis in Mice by Inhibiting Macrophage Apoptosis and Activation of the NLRP3 Inflammasome

doi: 10.1155/2021/6370911

Figure Lengend Snippet: Expression of IL-38 and its receptor Il-36R in mouse macrophages treated with LPS. (a) IL-38 normalized mRNA expression in mouse macrophages stimulated with LPS at the indicated concentrations for 12, 24, and 48 h. β -Actin served as the housekeeping gene. (b) IL-38 normalized mRNA expression in mouse macrophages stimulated with 1 μ g/ml LPS for 12, 24, or 48 h. β -Actin was used as the housekeeping gene. n = 3; ∗ P < 0.05 and ∗∗ P < 0.01 vs. the normal control (NC) group. NC was PBS treatment. (c) Serum concentrations of IL-38 in mice at the indicated timepoints after cecal ligation and puncture (CLP), based on ELISAs. n = 6; ∗∗ P < 0.01 vs. the sham group; (d) IL-36R normalized mRNA expression in mouse macrophages stimulated with LPS (1 μ g/ml; 24 h). β -Actin was used as housekeeping gene. (e, f) Representative western blot image (left panels) and quantification (right panels) of IL-38 and IL-36R protein levels of macrophages stimulated with 1 μ g/mL LPS for 24 h. β -Actin protein levels were used for normalization. (g, h) Representative immunofluorescence images of IL-38 (green) and IL-36R (green) of macrophages treated as in panels (d, e). Nuclei were stained with DAPI (blue). Magnification, ×600. Scale bar, 25 μ m. ∗∗ P < 0.01 vs . the NC group. Results are shown for three independent experiments.

Article Snippet: For in vivo blockade of IL-38, rat polyclonal anti-mouse IL-38 antibody (50 μ g per mouse; R&D Systems, Minneapolis, MN, USA) was injected intraperitoneally immediately after CLP, and a booster dose (50 μ g) was injected 24 h later.

Techniques: Expressing, Ligation, Western Blot, Immunofluorescence, Staining

Journal: Mediators of Inflammation

Article Title: IL-38 Alleviates Inflammation in Sepsis in Mice by Inhibiting Macrophage Apoptosis and Activation of the NLRP3 Inflammasome

doi: 10.1155/2021/6370911

Figure Lengend Snippet: Effects of IL-38 on macrophage functions under LPS stimulation. (a) Phagocytic activity of peritoneal mouse macrophages upon LPS stimulation (1 μ g/mL) and treatment with IL-38 at the indicated concentrations and time. Phagocytic activity was measured using the OD540 test. Phagocytic activity of macrophages treated with LPS (1 μ g/mL) and IL-38 (200 ng/mL) for 24 h in the right panel. Shown are arbitrary numbers. (b) Killing of L1210 cells by macrophages treated as indicated. Shown are arbitrary numbers. Killing activity of macrophages treated with LPS (1 μ g/mL) and IL-38 (200 ng/mL) for 24 h in the right panel. ∗ P < 0.05 and ∗∗ P < 0.01, vs . the NC group. NC was PBS treatment. n = 6 independent experiments. (c) Representative flow cytometry plots of macrophages treated for 24 h with PBS or LPS (1 μ g/mL) alone or with recombinant IL-38 (200 ng/mL). The M1 phenotype was F4/80 + CD11b + , and the M2 phenotype was F4/80 + CD206 + . Quantification of the experiment shown in the panel. The normal control (NC) group was PBS treatment. ∗ P < 0.05 and ∗∗ P < 0.01 vs . the NC group. ## P < 0.01 vs . the LPS group. n = 6 independent experiments.

Article Snippet: For in vivo blockade of IL-38, rat polyclonal anti-mouse IL-38 antibody (50 μ g per mouse; R&D Systems, Minneapolis, MN, USA) was injected intraperitoneally immediately after CLP, and a booster dose (50 μ g) was injected 24 h later.

Techniques: Activity Assay, Flow Cytometry, Recombinant

Journal: Mediators of Inflammation

Article Title: IL-38 Alleviates Inflammation in Sepsis in Mice by Inhibiting Macrophage Apoptosis and Activation of the NLRP3 Inflammasome

doi: 10.1155/2021/6370911

Figure Lengend Snippet: IL-38 dampens LPS-induced activation of the NLRP3 inflammasome in macrophages. (a) Representative western blot image of indicated proteins in macrophages stimulated with LPS (1 μ g/mL; 24 h) and/or IL-38 (200 ng/mL; 24 h). β -Actin protein was used for normalization. (b) Quantification of three independent experiments as shown in panel. (c) Representative immunofluorescence images of NLRP3 (green) of macrophages treated as in (a). Nuclei were stained with DAPI (blue). Magnification, ×600. Scale bar = 25 μ m. ∗ P < 0.05 and ∗∗ P < 0.01 vs . the normal control (NC) group. NC was PBS treatment. ## P < 0.01 vs . the LPS group. n = 3 independent experiments.

Article Snippet: For in vivo blockade of IL-38, rat polyclonal anti-mouse IL-38 antibody (50 μ g per mouse; R&D Systems, Minneapolis, MN, USA) was injected intraperitoneally immediately after CLP, and a booster dose (50 μ g) was injected 24 h later.

Techniques: Activation Assay, Western Blot, Immunofluorescence, Staining

Journal: Mediators of Inflammation

Article Title: IL-38 Alleviates Inflammation in Sepsis in Mice by Inhibiting Macrophage Apoptosis and Activation of the NLRP3 Inflammasome

doi: 10.1155/2021/6370911

Figure Lengend Snippet: IL-38 inhibits LPS-induced apoptosis of macrophages. (a) Representative flow cytometry plots of macrophages treated with PBS or LPS (1 μ g/mL; 24 h) alone or with recombinant IL-38 (200 ng/mL; 24 h) in order to evaluate apoptosis (left panel). Percentages of apoptotic (Annexin V+) macrophages (right panel). (b) Representative western blot image (left panels) and quantification (right panels) of the indicated proteins in macrophages stimulated with LPS (1 μ g/mL; 24 h) and/or IL-38 (200 ng/ml; 24 h). β -Actin protein was used for normalization. Normal control (NC) group was PBS treatment. ∗ P < 0.05 and ∗∗ P < 0.01, vs. the NC group; # P < 0 05 and ## P < 0.01, vs. the LPS group. n = 3 independent experiments.

Article Snippet: For in vivo blockade of IL-38, rat polyclonal anti-mouse IL-38 antibody (50 μ g per mouse; R&D Systems, Minneapolis, MN, USA) was injected intraperitoneally immediately after CLP, and a booster dose (50 μ g) was injected 24 h later.

Techniques: Flow Cytometry, Recombinant, Western Blot

Journal: Mediators of Inflammation

Article Title: IL-38 Alleviates Inflammation in Sepsis in Mice by Inhibiting Macrophage Apoptosis and Activation of the NLRP3 Inflammasome

doi: 10.1155/2021/6370911

Figure Lengend Snippet: IL-38 downregulation impacts polarization and inflammasome activation and apoptosis in LPS-treated macrophages. (a) Representative western blot image (left panels) and quantification (right panels) of indicated proteins in macrophages treated as indicated. β -Actin protein levels were used for normalization. (b) Representative flow cytometry plots of macrophages treated with PBS or LPS (1 μ g/mL; 24 h) alone or with anti-IL-38 siRNA in order to quantify populations with the M1 phenotype (F4/80 + CD11b + ) or M2 phenotype (F4/80 + CD206 + ). (c) Representative western blot image (upper panels) and quantification (lower panels) of indicated proteins in macrophages treated as indicated. β -Actin protein levels were used for normalization. (d) Representative immunofluorescence images of NLRP3 (green) of macrophages treated as indicated. Nuclei were stained with DAPI (blue). Magnification, ×600. Scale bar = 25 μ m. (e, f) Representative flow cytometry plots of macrophages treated with PBS or LPS (1 μ g/mL; 24 h) alone or with recombinant IL-38 (200 ng/mL; 24 h) in order to evaluate apoptosis (left panel). Percentages of apoptotic (Annexin V+) macrophages treated as indicated (right panel). Representative western blot image (left panels) and quantification (right panels) of the indicated proteins in macrophages stimulated with LPS (1 μ g/mL; 24 h). β -Actin protein was used for normalization. ∗ P < 0.05, vs. the Ctr siRNA group. # P < 0.05 and ## P < 0.01, vs. the LPS+Ctr siRNA group. Results reflect three independent experiments.

Article Snippet: For in vivo blockade of IL-38, rat polyclonal anti-mouse IL-38 antibody (50 μ g per mouse; R&D Systems, Minneapolis, MN, USA) was injected intraperitoneally immediately after CLP, and a booster dose (50 μ g) was injected 24 h later.

Techniques: Activation Assay, Western Blot, Flow Cytometry, Immunofluorescence, Staining, Recombinant

Journal: Mediators of Inflammation

Article Title: IL-38 Alleviates Inflammation in Sepsis in Mice by Inhibiting Macrophage Apoptosis and Activation of the NLRP3 Inflammasome

doi: 10.1155/2021/6370911

Figure Lengend Snippet: Effects of IL-38 stimulation or blockade on 7-day survival of septic mice. Kaplan Meier survival plot of mice treated as indicated. Septic mice were given PBS (blue line) or IL-38 (1 μ g per mouse), then subjected 2 h later to cecal ligation and puncture (CLP) (green line). Other animals were subjected to CLP, then immediately injected intraperitoneally with anti-IL-38 antibody (50 μ g per animal), followed 24 h later by a booster dose of 50 μ g (red line). ∗ P < 0.05 vs. the CLP+PBS group. Results are shown for 25 mice per group.

Article Snippet: For in vivo blockade of IL-38, rat polyclonal anti-mouse IL-38 antibody (50 μ g per mouse; R&D Systems, Minneapolis, MN, USA) was injected intraperitoneally immediately after CLP, and a booster dose (50 μ g) was injected 24 h later.

Techniques: Ligation, Injection

Journal: Mediators of Inflammation

Article Title: IL-38 Alleviates Inflammation in Sepsis in Mice by Inhibiting Macrophage Apoptosis and Activation of the NLRP3 Inflammasome

doi: 10.1155/2021/6370911

Figure Lengend Snippet: Effects of IL-38 on CLP-induced inflammation and markers of vital organ injury in septic mice. (a–g) Serum levels of indicated inflammatory biomarkers in CLP mice treated with IL-38 or anti-IL-38 antibody. ∗∗ P < 0.01 vs. the sham group; ## P < 0.01 vs. CLP control. (h) Mice that underwent CLP were treated with IL-38 or PBS. Mice were sacrificed 24 h after CLP. H&E staining and histological scores of the lungs, kidneys, and liver. ## P < 0.01 vs. the PBS group. Results are shown for six mice per group. # P < 0.05 and ## P < 0.01 vs. PBS control.

Article Snippet: For in vivo blockade of IL-38, rat polyclonal anti-mouse IL-38 antibody (50 μ g per mouse; R&D Systems, Minneapolis, MN, USA) was injected intraperitoneally immediately after CLP, and a booster dose (50 μ g) was injected 24 h later.

Techniques: Staining