biostudies-arrayexpressNikoletta KalenderoglouMus musculusNovogene Softwarehttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15152The experiment was designed to investigate the sequence of events controlling brown adipocyte differentiation, providing insight into brown fat physiology and metabolic dysfunction. Immortalized brown pre-adipocytes were treated with miRCURY miRNA Inhibitor that silenced miR-10b-5p and subjected cells to standard brown adipogenic treatment. Transcriptomic analysis was performed at days 0, 5, and 8 to identify pathways affected by miR-10b knockdown.biostudies-arrayexpressSample Treatment - Transfection: Adipocytes were plated in twelve-well plates at densities ranging between 200 and 250 K cells/well in media without antibiotics and transfected with either 50 nM miRNA inhibitors using RNAiMAX (Invitrogen). After 48 hours, adipocyte differentiation was initiated. Adipocyte differentiation: Plates were transferred to the 37 °C incubator and differentiation was initiated. Induction medium was administered for two days to initiate differentiation. The induction medium consisted of 1 µg/mL insulin, 1 nM 3,3′,5-triiodo-L-thyronine sodium salt (T3), 0.5 mM IBMX, 250 nM dexamethasone, and 125 µM indomethacin. After two days, the induction medium was replaced with maintenance medium, which contained 1 µg/mL insulin and 1 nM T3. Cells were maintained in this medium for the remainder of the experiment, with media changes performed as necessary.Sample Collection - Cultured cells were lysed in TRI reagent (Sigma).Nucleic Acid Extraction - RNA was isolated using TRI reagent.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).Growth Protocol - Immortalised brown and white adipocyte cell lines were generated by culturing the cells from the SVF of iBAT and scWAT of 10-week-old female 129Sv mice with retroviral-mediated expression of temperature sensitive simian virus 40 (SV40) large T-antigen (H-2Kb-tsA58. Cells were cultured in DMEM/F12 medium (Sigma) supplemented with 10% FBS Serum 10500 (Gibco), 1% L-Glutamine and 1% Penicillin-Streptomycin antibiotics (Gibco).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 FilesUnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq 6000RNA-seq of coding RNAMus musculusMark ChristianNikoletta KalenderogloufalseRNA Sequencing Analysis of miR-10b-5p Knockdown During Brown AdipogenesisThe experiment was designed to investigate the sequence of events controlling brown adipocyte differentiation, providing insight into brown fat physiology and metabolic dysfunction. Immortalized brown pre-adipocytes were treated with miRCURY miRNA Inhibitor that silenced miR-10b-5p and subjected cells to standard brown adipogenic treatment. Transcriptomic analysis was performed at days 0, 5, and 8 to identify pathways affected by miR-10b knockdown.2026-05-01T00:00:00Z2026-05-01T01:03:21.016Z2025-05-19T20:48:31.913ZE-MTAB-15152ERP172732EFO_0002944EFO_0004170EFO_0003789EFO_0005518EFO_0003738EFO_0003969EFO_0004184