title: miR-10b+/+.Day12.2
organism: Mus musculus
description: Protocols: Cultured cells were lysed in TRI reagent (Sigma). 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. 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. 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). 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).
attributes.tag: ENA first public
attributes.value: 2026-04-03
attributes.tag: INSDC center name
attributes.value: NTU
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-15151:miR-10b+/+.Day12.2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: mouse embryonic stem cells
attributes.tag: cell type
attributes.value: adipocyte differentiation day 12
attributes.tag: collection date
attributes.value: not collected
attributes.tag: common name
attributes.value: house mouse
attributes.tag: differentiation stage
attributes.value: adipogenic treatment day 12
attributes.tag: genotype
attributes.value: miR-10b+/+
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: progenitor cell type
attributes.value: embryonic stem cell
attributes.tag: sample name
attributes.value: E-MTAB-15151:miR-10b+/+.Day12.2
attributes.tag: scientific_name
attributes.value: Mus musculus