Pe cy7 anti crth2

ILC1s were sorted from PBMCs using a BD FACSAria. ILC1s were identified as Lin cocktail⁻(CD3, CD14, CD19, and CD20), CD34⁻, CD94⁻, TCRα/β⁻, TCRγ/δ⁻, CD1a⁻, CD45⁺, CD127⁺, CRTH2⁻, and CD117⁻ cells^{17 ,18 (link)}. FITC anti-Lin (BD Bioscience, USA), FITC anti-CD94, CD34, CD1a, TCRα/β, TCRγ/δ, APC-H7 anti-CD45, Percp-cy5.5 anti-CD127, PE-Cy7 anti-CRTH2, PE anti-CD117 (Biolegend, USA). Purity was routinely >99%. Cell viability, as determined by trypan blue staining, was >99% after isolation.

For further purification, MNCs from freshly resected patient tissue specimens were subjected to Ficoll‐Hypaque gradient centrifugation for 30 min at 24°C. Next, the MNC layer was transferred to a new tube, washed twice with phosphate-buffered saline (PBS), and suspended in PBS. ILCs were sorted using a BD FACSAria system (BD Bioscience) as Lin⁻-enriched MNCs as Lin cocktail⁻ (CD3, CD19, CD20, and CD14), CD94⁻ CD34⁻ CD1a⁻ TCRα/β⁻ TCRγ/δ⁻ CD45⁺ CD127⁺ CRTH2^+/− CD117^+/− cells using FITC anti-Lin (643510; BD Bioscience), CD94 (305504; Biolegend, San Diego, CA, USA), CD34 (343504; Biolegend), CD1a (300104; Biolegend), T cell receptor (TCR)α/β (306706; Biolegend), TCRγ/δ (331208; Biolegend), allophycocyanin (APC)-H7 anti-CD45 (56017; Biolegend), Percp-cy5.5 anti-CD127 (351322; Biolegend), phycoerythrin (PE)-Cy7 anti-CRTH2 (350118; Biolegend), and BV605 anti-CD117 (562687; Biolegend). pDCs were sorted as Lin⁻ CD94⁻ CD34⁻ CD1a⁻ TCRα/β⁻ TCRγ/δ⁻ CD45⁺ BDCA2⁺ cells using FITC anti-Lin, CD94, CD34, CD1a, TCRα/β, TCRγ/δ, APC-H7 anti-CD45, and APC anti-BDCA2 (17-9818-42; Biolegend). Purity was routinely >99%. Cell viability was determined by trypan blue staining and was >99% after isolation.