Pe cy7 conjugated anti mouse nk1

APC-conjugated anti-mouse TLR3 (11F8), APC-cy7-conjugated anti-mouse CD3 (145-2C11), FITC-conjugated anti-mouse CD3 (17A2), PerCP-cy5.5-conjugated anti-mouse CD4 (RM4–5), PE-conjugated anti-mouse CD8 (53–6.7), BV510-conjugated anti-mouse γδ TCR (GL3), PE-cy5-conjugated anti-mouse CD19 (6D5), PE-cy7-conjugated anti-mouse NK1.1 (PK136), APC-cy7-conjugated anti-mouse CD11b (M1/70), PE-conjugated anti-mouse Ly6G (1A8), PE-cy7-conjugated anti-mouse F4/80 (EMR1, Ly71), PE-conjugated anti-mouse Gr1(RB6-8C5), APC-conjugated anti-mouse CD69 (H1.2F3), PE conjugated anti-mouse Ly6G (1A8-Ly6g), PerCP-Cy5.5-conjugated anti-mouse CD11c (HL3), PE-anti-mouse CD25 (BC96), APC-anti-mouse CD69 (H1.2F3), FITC-conjugated anti-mouse CD94 (20d5), PE-conjugated anti-mouse CD314 (XMG1.2) were purchased from BioLegend (San Diego, CA, USA) and BD Pharmingen (San Diego, CA, USA).

For T lymphocyte subpopulations assay, single-cell suspension of splenocytes and mesenteric lymph nodes was adjusted to a concentration of 1 × 10⁶ cells/mL. T-cell specific markers were stained using PerCP-Cy5.5-conjugated anti-mouse CD3, FITC-conjugated anti-mouse CD4, Pacific Blue-conjugated anti-mouse CD8a, PE-conjugated anti-mouse CD25, PE/Cy7-conjugated anti-mouse NK-1.1 (only in splenocyte) and Alexa Fluor 647-conjugated anti-mouse foxP3 (only in mesenteric lymph node lymphocytes) (BioLegend, San Diego, CA, USA). We then conducted fluorescence activated cell sorting (FACS) and data analysis using a Beckman Gallios™ flow cytometer (Beckman Coulter, Pasadena, CA, USA), after adjusting the instrument settings using the control cells stained with isotype-matched antibodies.