GSM8449688: Neonatal ALI, Mock, rep1; Homo sapiens; RNA-Seq
For one 24-well ALI culture insert, the membrane was removed from the insert and treated with 500 µl of Accutase (Cat#A1110501, Thermo Fisher Scientific) with EDTA and EGTA (5 mM each) for 30 minutes at 37℃ on a sharker. After treatment, cells were dissociated by titrating and cell pellets were collected by centrifugation. After 1X PBS wash to remove the remaining Accutase, cells were resuspended in nuclear extraction buffer according to manufacturer's instruction (10X Genomics, protocol CG000124). The RNA library was constructed using 10X Single Cell 3' GEX and libraries were sequenced on Illumina NovaSeqS4 (150x150 Paired-End). single nuclei RNA sequencing