ATAC_ME3_WT_i8-day0_b
Mus musculus
Protocols: ME3 beige-like preadipocytes were grown to confluency and induced for adipogenic differentiation 2 days post confluency (day 0) by addition of adipogenic cocktail. On days 0, 1 and 7 of differentiation, cells were trypsinized, counted and 100,000 cells were slowly frozen down at -80C in culture medium containing 5% DMSO. Frozen cells were shipped on dry ice to Active Motif for processing. Cells were grown in AmnioMAX-C100medium supplementedwith 7.5% FBS, 7.5% C100 (all from Thermo Fisher Scientific,Waltham, MA, USA), 1% penicillin-streptomycin (PEST) (Sigma, St.Louis, MO, USA) and 2 mM L-glutamine (Sigma) at 37 °C and 5% CO2.Adipogenic differentiation was initiated three days post confluency(day 0) by induction medium containing 5 μg/mL Insulin (INS)(Sigma), 1 μMDexamethasone (DEX) (Sigma), 0.5mMisobutylmethylxanthine(IBMX) (Sigma) and 1 μM Rosiglitazone (ROSI) (CaymanChemical, Ann Arbor, MI, USA). From day 2 to day 4 only insulin wasadded to the basal medium and from day 4 to 7 cells were grown inthe basal medium. In-house protocols by Active Motif, based on published methods: Buenrostro, J.D., Giresi, P.G., Zaba, L.C., Chang, H.Y., Greenleaf, W.J., 2013. Transposition of native chromatin for fast and sensitive epigenomic profiling of open chromatin, DNA-binding proteins and nucleosome position. Nature Methods 10(12): 1213–8. Performed by in-house protocols by Active Motif, based on published methods: Buenrostro, J.D., Giresi, P.G., Zaba, L.C., Chang, H.Y., Greenleaf, W.J., 2013. Transposition of native chromatin for fast and sensitive epigenomic profiling of open chromatin, DNA-binding proteins and nucleosome position. Nature Methods 10(12): 1213–8.