Pe rat anti mouse cd86

Flow cytometry was performed on a CytoFLEX (Beckman), and data were analyzed by FlowJo software. The live cells were observed by the Fixable Viability Stain 780 (Cat#565388, BD Pharmingen). For cell-surface staining, cells were stained with BV510 Rat Anti-Mouse CD45(Cat#563891, BD Pharmingen), FITC Hamster Anti-Mouse CD3e (Cat#553061, BD Pharmingen), PE-Cy7 Rat Anti-Mouse CD4 (Cat#552775, BD Pharmingen), BV421 Rat Anti-Mouse CD25 (Cat#562606, BD Pharmingen), FITC Rat Anti-CD11b (Cat#557396, BD Pharmingen), BV421 Rat Anti-Mouse F4/80 (Cat#565411, BD Pharmingen), PE Rat Anti-Mouse CD86(Cat#561963 BD Pharmingen), Alexa Fluor 647 Rat Anti-Mouse CD206 (Cat#565250 BD Pharmingen). For intracellular staining, cells were stained with PE Rat Anti-Mouse IL-17A (Cat#561020 BD Pharmingen), Alexa Fluor 647 Rat anti-Mouse Foxp3(Cat#560402 BD Pharmingen). According to the reagent vendor’s instructions, Purified Rat Anti-Mouse CD16/CD32 is used to exclude background fluorescent signal interference, Cells fixation and permeabilization Fixation/Permeablization Kit (Cat#554714 BD Pharmingen) and Transcription Factor Buffer Set (Cat#562574 BD Pharmingen) were used for cell fixation and permeabilization.

Concanavalin A (Con A) and lipopolysaccharide (LPS) were purchased from ApexBio Technology Co., Ltd. and Sigma-Aldrich (Shanghai) Trading Co., Ltd., respectively. Mouse ELISA kits for IL-4 (20220111-20186A), IFN-γ (20220111-20140A), TNF-α (20220111-20852A), MCP-1 (20220111-20248A), IL-1 β (20220111-20174A), and IL-12 (20220111-20166A) were purchased from Shanghai Enzyme Linked Biotechnology Co., Ltd. The following flow cytometry antibodies were purchased: Brilliant Violet 605™ anti-Mouse CD45 (BioLegend), BV421 Rat anti-mouse CD25 (BD); BV510 Hamster anti-mouse CD3e (BD); mCD8-R208-APC (Sino Biological); mCD4-R711-FITC (Sino Biological); FITC anti-Mouse MHC-II (I-A/I-E) (eBioscience); BV421 Hamster anti-Mouse CD80 (BD); PE Rat anti-Mouse CD86 (BD); and BV786 Hamster ant-mouse CD11c (BD).