Biomarkers after radiation injury in an organ on a chip liver quad culture

Primary human hepatocytes, primary human LSECs, Kupffer cells and hepatic stellate cells were used using Emulate's proprietary orb and Zoe.Cells and liver chips were irradiated using an X Rad 320, which produces 320 kV photons at 12.5 mA. Cells were sham irradiated 0 Gy or received a single dose of 4 Gy or 8 Gy.Cell lysis for RNA isolation was performed as outlined by Emulate protocol. Raw reads from the RNAseq data were first filtered to remove reads with less than 10 percent uncertain nucleotides and/or greater than 50 percent proportion of low quality bases. Reads with adapter contamination were also removed.RNA sample quality was assessed by BioAnalyzer High Sensitivity RNA Assay and quantified by AccuBlue Broad Range RNA Quantitation assay. Equimolar pooling of libraries was performed based on QC values and sequenced on an Illumina NovaSeq 6000 with a read length configuration of 150 PE for 40 M PE reads per sample.