title: GSM5100070: BAL - Subject_2_3; Homo sapiens; RNA-Seq
library_construction_protocol: Bronchoscopy and BAL were performed as described previously (Mwandumba HC et al., J Immunol 2004; 172:4592-4598; Jambo KC et al., Thorax 2011; 66:375-382). A cell suspension containing 5 x106 BAC was centrifuged at 500g for 8 min at 4oC, the supernatant was discarded, and the cells were resuspended in 1mL of chilled 90% methanol (Merck Life Science, Dorset, UK). Fixed BAL samples were equilibrated on ice for 15', washed twice in rehydration buffer to remove residual methanol, stained with different HTO antibodies for 30' at 4C (as described above), and washed twice in cell staining buffer + 0.5U/ul of Rnase inhibitor (Sigma). Subsequently, the amount of cells recovered from each sample has been quantified and different samples were mixed together in a 1:1 ratio and multiplexed into the same 10X run. Libraries were generated using the same modifications described for mouse scRNA-seq samples.