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2 protocols using pe rat anti mouse f4 80

1

Isolation and Characterization of Murine Immune Cells

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Chicken egg ovalbumin (OVA, premium quality Grade V) was obtained from Sigma-Aldrich (St. Louis, MO). A TRPV4 KO mouse line on a C57BL/6 background was generated by Dr. Makoto Suzuki (Jichi Medical University, Tochigi, Japan) as described previously [22 (link)]. Inbred C57BL/6 mice (six-eight-week old) were obtained from Charles River (Wilmington, MA). Both male and female WT and TRPV4 KO mice were used in the experiments. The Institutional Animal Care and Use Committee (IACUC) of the University of Maryland reviewed and approved all animal research pertaining to this project, and all the animals were housed in HEPA filtered animal cages and provided ad libitum access to food and water.
APC-Cy7 rat anti-mouse CD45, APC-Cy7 rat IgG2b, FITC hamster anti-mouse CD11c, FITC Ar hamster IgG1, PE rat anti-mouse siglec-F, PE-rat IgG2a, APC-Cy7 rat anti-mouse Ly6G and Ly6C (Gr1), and APC-Cy7 rat IgG2b were purchased from BD Pharmingen (San Diego, CA); APC anti-mouse/human CD11b, APC rat IgG2b, and PE rat anti-mouse F4/80 were purchased from Biolegend (San Diego, CA).
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2

Inflammatory Signaling Pathway Characterization

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The kits for alanine aminotransferase (ALT), aspartate aminotransferase (AST), and triglyceride (TG) were supplied by Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Enzyme-linked immunosorbent assay (ELISA) kits for TNF-α and IL-1β were purchased from Bender MedSystems (Vienna, Austria). PE rat anti-mouse Ly6G was obtained from BD Biosciences (New Jersey, USA). Neutrophils and F4/80 antibodies were obtained from Thermo Scientific (Rockford, IL, USA). Rabbit anti-mouse TLR4 antibody, rabbit anti-mouse phospho-IRAK1, phospho-p38, phosphor-IKKβ, IKKβ, phospho-IκBα, IκBα, phospho-p65, p38, p65, β-actin, Lamin B1, and GAPDH were purchased from Abcam (Cambridge, UK). PE rat anti-mouse F4/80, FITC rat anti-mouse CD11b, and FITC rat anti-mouse CD45 antibodies were from Biolegend, Inc. (San Diego, USA). TLR4-specific antagonist TAK-242 was from MedChemExpress LLC (Shanghai, China). Palmitic acid (PA) was from Sigma-Aldrich (St. Louis, USA).
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