ChIP-seq in Azin2-/- and wt adipocyte progenitors

AZIN2 regulates adipogenesis and acetyl-coA levels in adipocyte progenitors. We investigated here how AZIN2 deficiency affects histone acetylation in adipocyte progenitors. Overall design: Adipocyte progenitors were isolated from subcutaneous adipose tissue of Azin2-/- and wild-type mice and cultured for 1 week 4.5 g/L glucose Glutamax–DMEM with 10% FBS and 1% penicillin/streptomycin at 37°C and 5% CO2. Then, they were cultured in medium containing 5 µg/ml insulin , 0.5 mM 3-isobutyl-1-methylxantine, 1 µM dexamethasone and 1 µM rosiglitazone. After 2 days, they were switched to medium containing 5 µg/ml insulin and cultured for another 5 days. Medium was changed every 2 days. ChIP was performed using the EZ-Magna ChIP HiSens (Millipore) following manufacturer's instructions. An antibody against H3K27Ac (Millipore 17- 683) was used. DNA was isolated and sequenced.