GSM7036418: Hypo24hr-3-1; Homo sapiens; RNA-Seq
To investigate the dynamic response of hISC to hypoxia at the whole transcriptomic level, we performed RNA-seq on human intestinal epithelium exposed to 6 hours, 24 hours, or 48 hours of hypoxia inside intestinal MPS, along with normoxic control intestinal MPS which were cultured for each respective time point with no hypoxia exposure. RNA samples were collected from each intestinal MPS, and Total RNA was extracted using RNAqueous-Micro Total RNA Isolation Kit (AM1931; ThermoFisher) according to manufacturer's protocols and stored at −80°C. Three technical replicates were collected from each duration of normoxia or hypoxia. RNA was extracted via the RNAqueous-Micro Total RNA Isolation Kit (AM1931; ThermoFisher, Waltham, MA) according to the manufacturer's protocol. To assess RNA quality prior to submission for sequencing, an RNA integrity number (RIN) was measured using the Agilent 2100 Bioanalyzer. After confirmation that each sample had a RIN of at least 8, integrated fluidic circuits (IFCs) for gene expression and genotyping analysis were prepared using the Advanta™ RNA-Seq NGS Library Prep Kit for the Standard Biotools Juno™. Bulk sequencing was run on a NovaSeq 6000 instrument using one lane of an S4 v1.5 flow cell (Illumina).