title: GSM8213910: 313491_1-Sample1_Mock; Homo sapiens; RNA-Seq
library_construction_protocol: Differentiated primary human bronchial epithelial cells (BEpC) were washed with PBS three times to remove mucus on the apical side of the insert. Basal media was removed  and basal side was washed once with PBS. Cells were then trypsinized by adding 0.25% Trypsin-EDTA (Thermo Fisher Scientific) on the basal and apical side of the insert for a total of 10 min at 37°C and resuspended in warm DMEM/F12 (Thermo Fisher Scientific) supplemented with 5% fetal bovine serum (FBS) (Thermo Fisher Scientific). Cells were centrifuged at 400*g (4°C) for 5 min and subsequently washed once in cold PBS supplemented with 0.04% bovine serum albumin (BSA) (Sigma-Aldrich). Finally, cells were resuspended in cold PBS + 0.04% BSA and filtered through a 40 μm Flowmi® filter (Merck) to produce a single cell suspension. The final cell concentration was around 600-1200 cells/ul, calculated with the Countess II Automated Cell Counter (ThermoFisher) using Trypan Blue Solution. The Gel Bead-in-emulsion (GEM) generation up to the 3' gene expression library construction was done following the 'Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 (Dual Index)' protocol (10x Genomics) and using the Chromium Controller (10x Genomics). Library was performed according to the manufacter's instructions (single cell 3' v3 protocol, 10x Genomics).