biostudies-arrayexpressHubert HacklHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-16150Bile acids (BAs), via the nuclear receptor FXR, regulate a complex transcriptional program to maintain fat, glucose, and protein metabolism. The mTORC1 signalling pathway integrates diverse nutrient/hormonal signals into FXR-related metabolic outputs, raising the question whether these transcriptional and posttranslational cascades are intertwined to maintain cellular homeostasis. Human HepG2 cells were treated with LXR agonist (GW3965), FXR agonist (GW4064) alone or combined with mTORC1 inhibitor Torin 1 (TOI), or combined with glucose/glutamine starved media (GLUST).biostudies-arrayexpressLibrary Construction - QuantSeq 3’ mRNA library preparation kit (Lexogen) according to manufacturer’s instructions.Growth Protocol - HepG2 cells were cultivated in DMEM (Gibco, 41965) containing 10% FBS (Gibco, 10500064) and 1% penicillin/streptomycin (Gibco, 15140122)Sample Collection - HepG2 cell line was ordered from ATCC (HB-8065) and cultured.Nucleic Acid Extraction - RNA was extracted using Trizol reagent (Invitrogen, 15596026) according to the manufacturer's instructions.Sample Treatment - Cells were treated with treated with GW3965 (5µM), GW4064 (5 µM) alone or combined with mTORC1 inhibitor Torin 1 (TOI), or combined with glucose/glutamine starved media (GLUST). DMSO with final concentration of 0.1% (v/v) was used as control treatment.Sequencing - RNA sequencing on samples from human HepG2 cell line was performed at the Next Generation Sequencing Core facility at the Medical University of Innsbruck on an Ion Torrent Proton device (ThermoFisher)OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesSequence Alignment - Trimmomatic (v 0.36) was used to cut single-end sequencing reads at a length of 200bp, remove adaptor sequences and sequencing reads with minor quality as assessed by FastQC. The splice-aware aligner STAR (v 2.5.3a) was used to map reads to the UCSC human reference genome (hg38) and RefSeq gene annotation with 200 bp splice junction overhangs. HTSeq (v 0.9.1) was used to quantify raw gene counts based on refGene annotation.Data Transformation - Counts were normalized using DESeq2, log2-transformed with adding a pseudocount of 1.MetabolomicsUnknownTranscriptomicsGenomicsProteomicsIon Torrent ProtonRNA-seq of coding RNAHomo sapiensHubert HacklTarek MoustafaAlex ZeufelfalseTranscriptomic analysis of nuclear receptor(LXR, FXR) modulation, mTOR inhibition, and nutrient stress on human HepG2 cellsBile acids (BAs), via the nuclear receptor FXR, regulate a complex transcriptional program to maintain fat, glucose, and protein metabolism. The mTORC1 signalling pathway integrates diverse nutrient/hormonal signals into FXR-related metabolic outputs, raising the question whether these transcriptional and posttranslational cascades are intertwined to maintain cellular homeostasis. Human HepG2 cells were treated with LXR agonist (GW3965), FXR agonist (GW4064) alone or combined with mTORC1 inhibitor Torin 1 (TOI), or combined with glucose/glutamine starved media (GLUST).2025-12-16T00:00:00Z2026-05-30T17:40:20.515Z2025-11-17T11:20:50.337ZE-MTAB-16150ERP185155EFO_0002944EFO_0004170EFO_0003789EFO_0004917EFO_0005518EFO_0003816EFO_0003738EFO_0004184EFO_0003969