Anti klrg1 2f1

For cell sorting and flow cytometry analysis, cell suspension were prepared as described above, incubated with fixable viability dye and subsequently stained with following fluorophore-conjugated monoclonal antibodies: anti-CD45 (30-F11, BD Biosciences), anti-CD45.2 (104, Biolegend), anti-CD90.2 (thy1.2) (53–2.1, Biolegend), anti-CD3 (17A2, Biolegend), anti-CD4 (Biolegend, RM4–5), anti-CD8 (53–6.7, Biolegend), anti-CD11b (M1/70; Biolegend), anti-CD11c (N418, Biolegend), anti-Ly6C (HK1.4, Biolegend), anti-Ly6G (1A8, BD Biosciences), anti-Siglec-F (E50–2440, BD Bioscience), anti-mouse MHC Class II (I-A/I-E) (M5/114.15.2, eBioscience), anti-NK1.1 (PK136, Biolegend), anti-CD19 (6D5, Biolegend), anti-T1/ST2 (DJ8, MD Biosciences), anti-KLRG1 (2F1, Biolegend), anti-FoxP3 (FJK-16S, eBiosciences), anti-Ki-67 (16A8, Biolegend), anti-Gata3 (TWAJ, eBioscience), anti-IL13 (eBio13A, eBioscience), anti-IFNgR1 (XMG1.2, BD Bioscience).

Tumors were dissociated using the MACs mouse tumor dissociation kit (Miltenyl Biotec) and suspended in PBS+1% FBS. The cells were incubated in blocking buffer containing 1:200 CD16/32(clone 93, eBioscience) for 30 min on ice. They were subsequently stained by fluorescent-conjugated primary antibodies at previously validated concentrations for 1 hr on ice. Following three PBS+1 % FBS washes, the cells were resuspended in ice cold PBS or fixed in 1 % PFA for immediate acquisition on the LSR Fortessa at the Baylor College of Medicine FACS and Cell Sorting core. Data was further analyzed using FlowJo Software version 10.0.
The following antibodies against mouse antigens were used: anti-CD3ε (145-2C11), anti-CD4 (GK1.5), anti- CD8 (53-6.7) (all from eBioscience); anti-B220 (RA3-6B2), anti-CD11b (M1/70)), anti-CD45 (30-F11), anti- F4/80 (BM8), anti-Ly6G(IA8)(all from Tonbo); anti-CD127 (SB/199, BD Biosciences), anti-CD44 (IM7, BD Biosciences), anti-CD62L (MEL14, Biolegend), Anti-KI67(16A8, Biolegend), anti-CSF1R(AFS98, Biolegend), anti-KLRG1(2F1, Biolegend).