MM3 dataset, mouse splenocytes

Splenocytes from C57Bl/6 mice were disrupted with glass slides, filtered through 100 μm mesh, and red blood cells lysed. Cells were stained with Fixable Viability Dye eFluor780 (eBioscience), anti-CD90.2–BV510 (1:250, clone 53-2.1), anti-CD25–BV650 (1:200, clone PC61), anti-CD45–BUV395 (1:500, clone 30-F11) (all Biolegend), anti-CD127–PE (1:100, clone A7R34) and anti-B220–PE-Cy5 (1:200, clone RA3-6B2) (all eBioscience). Cells were fixed and permeabilized with Foxp3 transcription factor staining buffer set (eBioscience) according to the manufacturer’s instructions, and stained overnight at 4 °C with the following antibodies: anti-T-bet–BV421 (1:200, clone 4B10), anti-CD8–BV785 (1:2000, clone 53-6.7), anti-NKp46–FITC (1:500, clone 29A1.4), anti-NK1.1–PE-Cy5.5 (1:2500, clone PK136), anti-MHCII–AF700 (1:2000, clone M5/114.15.2) (all Biolegend), anti-CD11b–eFluor450 (1:1000, clone M1/70), anti-GATA3–PE-Cy7 (1:100, clone L50-823), anti-CD3–biotin (1:1000, clone 145-2C11), anti-RORt–APC (1:500, clone AFKJS-9) (all eBioscience), anti-TCRβ–BB790-P (1:4000, clone H57-597), anti-CD4–BUV496 (1:500, clone GK1.5), and anti-CD19–BUV661 (1:2000, clone 1D3) (all BD Biosciences). Antibodies used for compensation controls were anti-CD25–BV421 (1:200, clone PC61), anti-CD44–BV510 (1:200, clone IM7), anti-CD3–BV650 (1:200, clone 17A2), anti-CD8–BV785 (1:2000, clone 53-6.7), anti-NK1.1–PE-Cy5.5 (1:2500, clone PK136), anti-MHCII–AF700 (1:2000, clone M5/114.15.2) (all Biolegend), anti-CD11b–eFluor450 (1:1000, clone M1/70), anti-TCRβ–FITC (1:500, clone H57-597), anti-B220–PE-Cy5 (1:200, clone RA3-6B2), anti-CD23–PE-Cy7 (1:500, clone B3B4), anti-CD8–biotin (1:200, clone 53-6.7), anti-Foxp3–APC (1:200, clone FJK-16s), anti-CD69–PE (1:200, clone H1.2F3) (all eBioscience), anti-TCRβ–BB790-P (1:4000, clone H57-597), anti-CD103–BUV395 (1:500, clone M290), anti-CD4–BUV496 (1:200, clone GK1.5), and anti-CD19–BUV661 (1:2000, clone 1D3) (all BD Biosciences). Streptavidin AF350 (1:200, Invitrogen) was used to identify biotinylated antibody. Samples were acquired on a Yeti/ZE5 flow cytometer (Propel Labs/BioRad).