RNA-Seq of Homo sapiens: flow-sorted macrophages from bronchoalveolar lavage fluid: Sample Sample_18
Samples were enriched by flow cytometry sorting for live cells, excluding granulocytes. Cells were sorted into 2% BSA in Dulbecco's PBS (DPBS), pelleted by centrifugation at 400g for 5 min at 4 °C, resuspended in 0.1% BSA in DPBS to about 1,000 cells μl−1. Concentration was confirmed using K2 Cellometer (Nexcelom) with AO/PI reagent and cells were loaded on 10x Genomics Chip A with Chromium Single Cell 5′ gel beads and reagents (10x Genomics) aiming to capture around 5,000–10,000 cells per library. Libraries were prepared according to the manufacturer's protocol (10x Genomics, CG000086_RevM). After quality checks, single-cell RNA-seq libraries were pooled and sequenced on a NovaSeq 6000 instrument.