characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-08-31T01:36:53Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-08-31T01:36:53Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA111346729
characteristics.characteristic: External Id
characteristics.text: KIS
characteristics.characteristic: INSDC center name
characteristics.text: 2023-08-31T01:36:53Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-08-31T01:36:53Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS13444278
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12160:NR-1 10nM 4h Bio1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: immortalized preadipocyte cell line
characteristics.characteristic: cell_line
characteristics.text: brown adipocyte
characteristics.characteristic: cell_type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Cells were washed with PBS and lysed directly in cell culture well using lysis buffer provided in Quick-RNA MicroPrep Plus kit (Zymo Research). Brown adipocytes were differentiated from an immortalized brown preadipocyte mouse cell line previously described (PMID: 16679291). Preadipocytes were maintained and expanded at low confluence (<50 %) in growth media [DMEM high glucose (Gibco) supplemented with 20% fetal bovine serum (Sigma-Aldrich), 20 mM HEPES (Sigma-Aldrich), and 100 U/mL penicillin /streptomycin (Gibco)]. For differentiation, preadipocytes were washed twice with phosphate-buffered solution (PBS), dissociated with TrypLE Express, and seeded at a density of approximately 23,000 cells/cm2 in growth medium. Medium was changed after 24 hours to differentiation medium [DMEM high glucose supplemented with 10% fetal bovine serum, 20 nM insulin, 1 nM triiodothyronine (T3), and 100 U/mL penicillin/streptomycin] and cells were cultured for 2 days. Cell differentiation was then induced by addition of induction media (differentiation media supplemented with 0.125 mM indomethacin, 0.5 μM dexamethasone, and 0.5 mM isobutyl methylxanthine) for two days, after which, medium was changed to differentiation medium for two additional days. Differentiated adipocytes were serum starved for 2 hours using starvation medium [DMEM low glucose (Gibco) supplemented with 2.5 mM glucose (to a final concentration of 8mM), 0.5% bovine serum albumin (BSA) (Sigma-Aldrich), and 100 U/mL penicillin /streptomycin] before treatment with Insulin-NanoRod structures or insulin diluted in starvation medium for four hours. For control sample only starvation media was used. Following treatment, cells were washed one time in PBS and harvested for RNA isolation. Total RNA was isolated from differentiated adipocytes using Quick-RNA MicroPrep Plus kit (Zymo Research) TruSeq RNA Library Prep Kit v2 Low Sample Protocol, 500ng RNA input
characteristics.characteristic: description
characteristics.text: neonate
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: interscapular brown adipocyte tissue
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: C57BL/6J
characteristics.characteristic: strain
characteristics.text: NR-1 10nM 4h Bio1
characteristics.characteristic: title