title: Sample 4
organism: Mus musculus
description: Protocols: Cells were washed in PBS, lysed by addition of RLT buffer (Qiagen, Hilden, Germany) and homogenized by centrifugation on QIAShredder columns (Qiagen) before snap-freezing in liquid nitrogen. Mouse embryonic fibroblast Rb-/- Line 3 (ME3) cells were previously generated (41) and characterized to be a model of beige adipocytes (42-46). Cells were grown in AmnioMAX -C100 (Thermo Fisher Scientific, Waltham, Massachusetts, USA) supplemented with 7.5% FBS (Thermo Fisher Scientific, Waltham, MA, USA ), 7.5% C100 (Thermo Fisher Scientific), 1% penicillin-streptomycin (PEST) (Sigma, St. Louis, MO, USA) and 2 mM L-glutamine (Sigma) at 37oC and 5% CO2. Cells were initiated to differentiate three days post confluency (day 0) by induction medium containing 5 ug/mL Insulin (INS) (Sigma), 1 μM Dexametasone (DEX) (Sigma), 0.5 mM isobutyl methylxanthine (IBMX) (Sigma) and 1 μM Rosiglitazone (ROSI) (Cayman Chemical, ann Arbor, MI, USA). From day 2 to day 4 only insulin was added to the basal medium and from day 4 to 7 cells were grown in the basal medium. RNA was isolated using the RNeasy Mini kit (Qiagen) with on-column DNaseI-treatment and RNA integrity was validated by the RNA 6000 Nano kit on the 2100 BioAnalyzer (Agilent, Santa Clara, USA). Library preparation and 2x75bp paired-end mRNA sequencing was performed using the TruSeq Stranded mRNA kit (Illumina, Sand Diego, CA, USA)
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-11-26T04:04:49Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-07-29T18:23:46Z
attributes.tag: External Id
attributes.value: SAMEA5817857
attributes.tag: INSDC center name
attributes.value: UNIVERSITY OF BERGEN
attributes.tag: INSDC first public
attributes.value: 2019-11-26T04:04:49Z
attributes.tag: INSDC last update
attributes.value: 2019-07-29T18:23:46Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8200:Sample 4
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: ME3
attributes.tag: cell type
attributes.value: mouse embryonic fibroblast cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: embryo
attributes.tag: disease
attributes.value: normal
attributes.tag: genotype
attributes.value: Rb homozygous knockout
attributes.tag: organism part
attributes.value: embryo
attributes.tag: sample name
attributes.value: E-MTAB-8200:Sample 4
attributes.tag: scientific_name
attributes.value: Mus musculus