Anti cd103

Cells were stained with the following antibodies obtained from BD Biosciences (Franklin Lakes, NJ, USA), eBioscience, or BioLegend (San Diego, CA, USA): anti-CD4 (BioLegend, 100451), anti-CXCR3 (eBioscience, 12-1831-82), anti-CCR7 (eBioscience, 12-1971-82), anti-CD44 (eBioscience, 12-0441-83), anti-CD62L (BioLegend, 104406), anti-CD11c (BD Bioscience, 553801), anti-MHCII (BioLegend, 107631), anti-CD80 (BD Biosciences, 553769), anti-CD86 (BD Biosciences, 553692), anti-B220 (eBioscience, 12-0452-83), anti-CD8 (BD Bioscience, 553032), anti-CD103 (BD Bioscience, 557495), anti-CD45 (BioLegend, 103132), and anti-CD11b (BioLegend, 101263). For Th1 and Treg analyses, cells were stained for surface markers, permeabilized with the Intracellular Fixation and Permeabilization Buffer Set (eBioscience, 88-8824-00), and then stained with anti-IFN-γ (BioLegend, 505825) and anti-FOXP3 (eBioscience, 17-5773-82) antibodies. The following antibodies were obtained from Cell Signaling Technology (Danvers, Massachusetts, USA): anti-Ezh2 (#5246), anti-Runx1 (#4336), and anti-H3K27me3 (#9733). The Alexa Fluor™ 488 Goat Anti-Rabbit SFX Kit from Invitrogen (Carlsbad, CA, USA) was used as a secondary antibody. Multicolor flow cytometric analysis was performed using a CytoFLEX LX (Beckman Coulter, Indianapolis, IN, USA).

Cell surface staining were performed with the following antibodies: anti-CD3ε, and anti-CD4, anti-CD11b, anti-CD11c, anti-CD19, anti-B220, and anti-Ly-6G (TONBO Bioscience, San Diego, CA, USA); anti-IL-33R (ST2), anti-TCRγδ, anti-CD90.2, and anti-CD45 (BioLegend, San Diego, CA, USA); anti-TCRβ, anti-CD25, anti-CD44, anti-CD62L, anti-CD69, anti-CD103, anti-NK1.1, and anti-TER119 (BD Bioscience); and anti-CD127 (eBioscience, San Diego, CA, USA). The same procedures were conducted with the following antibodies for the intracellular staining: anti-IL-4 and anti-IL-5 (BD Bioscience); and anti-IL-13 (eBioscience). Fixable Viability Dye (eBioscience) was used to exclude dead cells. All samples were analyzed using the FACS Verse (BD Bioscience) and Flow Jo software programs (Tree Star Inc., San Carlos, CA, USA).

Cells were suspended in PBS supplemented with 10% FBS and 0.02% sodium azide for cell surface staining. Cells were stained with: anti-CD103, anti-CD11c, anti-CD86 (BD Biosciences, Pharmingen, San Diego, CA, USA), anti-CD90.2, anti-CD4, anti-CD8α, anti-MHC-II, anti-XCR1 (BioLegend, San Diego, CA, USA), anti-NK1.1, anti-CD19 (Ablab, Vancouver, B.C., Canada), anti-CD11b (eBioscience, San Diego, CA, USA). For intracellular staining, cells were fixed and permeabilized using the Intracellular Fixation & Permeabilization Buffer Set (eBioscience). Antibodies used were: anti-IL-13 (eBioscience), anti-IFNg, and anti-IL-17A (BioLegend).