3T3-L1 cells were transfected using the LipofectAMINE RNAimax reagent (Invitrogen) according to the manufacturerÆs protocol. Final concentration of siRNA was 50nM. SMARTpool siRNA and negative control siRNA (siCONTROL non-targeting pooled, Dharmacon) were used. Transfected 3T3-L1 fibroblasts were differentiated into adipocytes by treating confluent cells with 0.5mM isobutylmethylxanthine (IBMX) and 0.25?M dexamethasone for 48hrs and then with insulin (5?g/ml) and 1?M Rosiglitazone for 48hrs. Media was replaced every 2 days. Cells were cultured and harvested on day -2, 0, 1, 2, 3, 4, 5, 6 days in relation to rosiglitazone treatment. Three independent biological replicates were collected.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted directly from the cells using the Qiagen RNeasy kit (Qiagen).
