{"database":"biostudies-arrayexpress","file_versions":[],"scores":null,"additional":{"submitter":["Nikoletta Kalenderoglou"],"organism":["Mus musculus"],"software":["Novogene Software"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/E-MTAB-15152"],"description":["The 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."],"repository":["biostudies-arrayexpress"],"sample_protocol":["Sample 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)."],"figure_sub":["Organization","MINSEQE Score","Assays and Data","MAGE-TAB Files"],"omics_type":["Unknown","Transcriptomics","Genomics","Proteomics"],"instrument_platform":["Illumina NovaSeq 6000"],"study_type":["RNA-seq of coding RNA"],"species":["Mus musculus"],"pubmed_authors":["Mark Christian","Nikoletta Kalenderoglou"],"additional_accession":[]},"is_claimable":false,"name":"RNA Sequencing Analysis of miR-10b-5p Knockdown During Brown Adipogenesis","description":"The 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.","dates":{"release":"2026-05-01T00:00:00Z","modification":"2026-05-01T01:03:21.016Z","creation":"2025-05-19T20:48:31.913Z"},"accession":"E-MTAB-15152","cross_references":{"ENA":["ERP172732"],"EFO":["EFO_0002944","EFO_0004170","EFO_0003789","EFO_0005518","EFO_0003738","EFO_0003969","EFO_0004184"]}}