biostudies-arrayexpressNikoletta KalenderoglouMus musculusNovogene Softwarehttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15151The experiment was designed to investigate the sequence of events regulating preadipocyte commitment and terminal differentiation into adipocytes, providing insight into brown fat physiology and metabolic dysfunction. miR-10b was found to be upregulated in brown adipocytes, and CRISPR/Cas9 was used to generate its functional knockout (KO) in E14 mouse embryonic stem cells (ES). Transcriptomic analysis was performed at days 0, 12, and 27 to identify pathways affected by miR-10b depletion.biostudies-arrayexpressNucleic Acid Extraction - RNA was isolated using TRI reagent.The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Library preparation was performed by Novogene (Novogene Co., Ltd).Sample Collection - Cultured cells were lysed in TRI reagent (Sigma).Growth Protocol - Mouse embryonic stem cells (mESCs) were cultured in Glasgow's MEM (GMEM) (Invitrogen) supplemented with 10% FBS, 2 mM L-Glutamine (Sigma), 0.5% Penicillin-Streptomycin (Gibco), 0.1mM MEM Non-Essential Amino Acids Solution (Gibco), 0.25% Sodium Bicarbonate solution (Gibco, 7.5% stock), 1mM Sodium Pyruvate (Gibco), 0.1mM 2-Mercaptoethanol (Gibco) and 1000units/ml LIF (Merckmillipore) on 0.1% gelatin (Millipore)-coated flasks.Sample Treatment - mESCs were induced to differentiate into mature adipocytes. Briefly, the cell suspension is prepared at 1 × 10⁵ cells/mL in differentiation medium. 20 μL drops are dispensed onto the inner lid of a 150 mm Petri dish, inverted over the dish, and incubated at 37°C with 5% CO₂ for 48 hours. Embryonic Bodies are collected and plated in a gelatin-precoated 6-well plate. On Day 3, the medium is replaced with 1 μM ATRA and 12.5 μg/mL Ascorbic Acid (AsA), refreshed daily. On Day 7, ATRA is removed, and the medium is replaced with 12.5 μg/mL AsA, 3 nM T3, 0.5 μg/mL insulin, and 0.5 μM rosiglitazone. On Day 12, cells are dissociated with accutase, resuspended in 2 mL differentiation medium, and replated. From Day 15, the medium is changed every 3 days, supplemented with 25 μg/mL AsA, 0.1 μM dexamethasone, 20 μg/mL insulin, 0.5 μM rosiglitazone, 0.06 mM indomethacin, and 0.5 μM IBMX. On Days 21 and 24, the medium is replaced with 25 μg/mL AsA, 20 μg/mL insulin, 0.5 μM rosiglitazone, and 3 nM T3. Adipocyte differentiation is assessed on Day 27 by the presence of lipid droplets.Library Construction - Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using dTTP for non-directional library. For the non-directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. Library preparation was performed by Novogene (Novogene Co., Ltd).Sequencing - PolyA library prep, PE150 sequencing, 30 million paired reads per sample. Library preparation and sequencing with Illumina technology were performed by Novogene (Novogene Co., Ltd).OrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesMetabolomicsUnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq 6000RNA-seq of total RNAMus musculusMark ChristianNikoletta KalenderogloufalseRNA Sequencing of miR-10b +/+ and miR-10b -/- Mouse Embryonic Stem Cells During Adipocyte Differentiation at Days 0, 12, and 27The experiment was designed to investigate the sequence of events regulating preadipocyte commitment and terminal differentiation into adipocytes, providing insight into brown fat physiology and metabolic dysfunction. miR-10b was found to be upregulated in brown adipocytes, and CRISPR/Cas9 was used to generate its functional knockout (KO) in E14 mouse embryonic stem cells (ES). Transcriptomic analysis was performed at days 0, 12, and 27 to identify pathways affected by miR-10b depletion.2026-05-01T00:00:00Z2026-05-01T01:03:27.384Z2025-06-09T14:26:32.551ZE-MTAB-15151ERP173140EFO_0002944EFO_0004170EFO_0009653EFO_0003789EFO_0005518EFO_0003969EFO_0004184