Anti pe antibody coated magnetic beads

MDSCs were isolated as described earlier (Chakraborty et al., 2017 (link)). Briefly, lungs were dissociated in collagenase-DNase suspension (Collagenase from Clostridium histolyticum, Cat # 10103586001, Roche and DNAase I from Bovine Pancreas, Cat # 11284932001, Roche) using a gentle MACS Dissociator (Miltenyi Biotec) according to the manufacturer’s protocol. Single-cell suspensions were obtained by passing the dissociated tissue through a 70 μm cell strainer (BD Biosciences) and washed with PBS containing 2% FBS. CD11b cells were enriched by labeling them with PE and using anti-PE antibody-coated magnetic beads (Cat # 130048801, Miltenyi Biotec). Live cells were gated from these enriched cells using a cell viability dye (SYTOX Green Ready Flow Reagent, Invitrogen, Cat# R37168) and then sorted to 90%–95% purity by flow cytometry (BD FACS Melody) using fluorochromelabeled antibodies to sort CD11b⁺ F4/80⁺ Ly6G^int Ly6C^lo cells.