biostudies-arrayexpressUnknownTranscriptomicsGenomicsProteomicsJames ClelandNextSeq 2000RNA-seq of coding RNAMus musculus domesticusMus musculus domesticushttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-17079To test the contribution of several sex-biased genes to hepatocyte transcription, we performed bulk RNA-sequencing on female primary hepatocytes.biostudies-arrayexpressSample Collection - Primary hepatocytes were isolated by ex vivo perfusion of liver left lateral lobes using the gentleMACS Octo Dissociator with Heaters and the associated Liver Perfusion Kit. Cells were cultured at 37°C and 5% CO2 for 3 h in collagen-coated 6-well plates containing DMEM supplemented with 5% FCS and 1% Pen Strep (Charni-Natan and Goldstein 2020), cultured overnight (for 14-21 h) in William’s E medium supplemented with 2 mM GlutaMAX and 1% Pen Strep (Charni-Natan and Goldstein 2020), cultured for 30 min in William’s E medium supplemented with 2 mM GlutaMAX (Charni-Natan and Goldstein 2020). siRNAs were delivered using Lipofectamine RNAiMAX Reagent according to the manufacturer's instructions (25 pmol siRNA and 7.5µl RNAiMAX per well). At 48 hpt, primary hepatocytes were incubated in 0.25% Trypsin-EDTA solution for 6 min at 37°C and 5% CO2, transferred to 2 ml screw cap micro tubes, rinsed once with PBS to dilute out residual proteins from the trypsin solution, then resuspended in 1 ml PBS. 20% of each sample was transferred to a new 1.5 ml DNA LoBind tube tube for downstream isolation of RNA and protein. Cells transfected with Luciferase siRNA or Negative Control No. 2 siRNA were used as negative controls.Sequencing - Libraries were sequenced on a NextSeq 2000 using P2 or P3 XLEAP-SBS 100 cycles kits.Nucleic Acid Extraction - RNA was extracted from 20% of the primary hepatocytes in each transfected 6-well plate well using a Quick-RNA Microprep Kit.Library Construction - Library preparation was performed using the NEBNext Ultra Express RNA Library Prep Kit and NEBNext Poly(A) mRNA Magnetic Isolation Module.OrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesJames ClelandfalseBulk RNA-seq of primary hepatocytes after knockdown of candidate sex-biased lipid metabolism regulatorsTo test the contribution of several sex-biased genes to hepatocyte transcription, we performed bulk RNA-sequencing on female primary hepatocytes.2026-06-11T00:00:00Z2026-06-11T01:00:44.168Z2026-05-26T12:26:59.155ZE-MTAB-17079ERP193965E-MTAB-13987EFO_0002944EFO_0004170EFO_0005518EFO_0003738EFO_0004184