GSM5770211: A2, WT-Air, 002, S40; Mus musculus; RNA-Seq
(Lin-Ckit+Sca1+) cells were enriched inside the hypoxia chamber using mouse lineage cell depletion kit , EasySep™ release mouse biotin positive selection kit and EasySep™ mouse SCA1 positive selection kit consecutively. LSK enriched cells were divided into two groups; group 1 was incubated inside the hypoxia chamber for RNA extraction, and group 2 was equilibrated in ambient air for 6 hours before RNA extraction. Total RNA was extracted using Qiagen RNeasy Plus Micro Kit. RNA with Agilent Bioanalyzer RIN of > 7 were selected for library preparation. Library preparation was performed using New England Biolabs Single Cell/Low Input RNA Library Prep Kit for Illumina. Paired-end RNA-sequencing (RNA-seq) was performed on Illumina instrument by IU School of Medicine Medical Genomics core. Each group/condition consists of four matched biological replicates and ~30 million reads were obtained per sample.