Pe hamster anti mouse cd11c

100 μL of sample, containing of 1 × 10⁶ cells, was first incubated with Fc receptor- blocking antibody (anti-CD16/CD32; BD Pharmingen, USA) for 5 min to reduce non-specific binding. Next, the sample was labeled for 20 min in the dark at 4 °C, with the following anti-CD primary antibodies: PE hamster anti-mouse CD11c (BD Pharmingen, USA), FITC rat anti-mouse CD86 (BD Pharmingen), APC anti-mouse MHC Class II (eBiosciences, USA). Labeled cells were washed three times with PBS supplemented with 2% bovine serum albumin (Sigma-Aldrich) and 0.1% NaN₃, and fixed. Flow cytometric analysis was performed on a Becton-Dickinson LSRII (USA).

Freshly harvested pancreatic and colonic tissues were digested in 1.0 mg/mL collagenase-P (Boehringer, Mannheim, Germany) solution at 37°C for 15 min and filtered through 75 µm filters with hank’s solution (Beyotime, Shanghai, China). Single-cell suspensions were incubated for 30 min at 4°C in hank’s solution with the following mAbs: APC Rat Anti-Mouse CD11b, BV421 Rat Anti-Mouse F4/80, Alexa Fluor 700 Rat Anti-Mouse Ly-6G, PE Hamster Anti-Mouse CD11c, FITC Rat Anti-Mouse MHCII, and PerCP-Cy5.5 Rat Anti-Mouse CD8a (BD Pharmingen, CA, USA). Gating method of fluorescence-activated cell sorting was programmed as CD11b⁺ Ly-6G⁺ (for neutrophils), CD11b⁺ F4/80⁺ (for macrophages), CD11c⁺ MHCII⁺ (for conventional dendritic cells, cDCs), and CD8a⁺CD11c⁺ MHCII⁺ (for plasmacytoid dendritic cells, pDCs). Flow cytometer was performed on Attune NxT (Thermo Fisher Scientific, MA, USA). Data were analyzed using ACEA NovoExpress software (Novo Express International, Inc., South San Francisco, CA, USA).

The phenotypes of DC as well as the proliferation assay of CFSE-labeled T cells were determined by flow cytometry^{35 (link)}. For cell surface staining, single-cell suspensions were incubated for 15 min at 4 °C with PE Hamster anti-mouse CD11c (0.4 μg/ml) (BD Biosciences, 553802), PerCP-Cy5.5 Hamster anti-mouse CD80 (0.2 μg/ml) (BD Biosciences, 560526), PerCP-Cy5.5 anti-mouse CD86 (0.2 μg/ml) (Biolegend, 105027), PE-Cy7 anti-mouse CD40 (0.1 μg/ml) (Biolegend, 124621), PE-Cy7 anti-mouse I-Ab (0.1 μg/ml) (Biolegend, 116420), PE-Cy7 anti-mouse CD4 (0.2 μg/ml) (Biolegend, 100422), APC anti-mouse TCR Vβ5.1 antibody (0.2 μg/ml) (Biolegend, 139511), and FITC anti-mouse CD45.2 antibody (0.2 μg/ml) (Biolegend, 109805). Samples were washed and analyzed by FACS versus flow cytometry (BD Biosciences), and the gating strategies were shown in Supplementary Fig. 9.