RNAseq analysis of 3T3-L1 adipocytes cultured in high and low media volumes
3T3-L1 fibroblasts were cultured in high glucose DMEM supplemented with 10% foetal bovine serum (FBS) and 2 mM glutamax at 37°C in 10% CO2. For differentiation into adipocytes, fibroblasts were seeded into 12-well plates, and cultured in 1 mL/well of 10% FBS-supplemented DMEM containing an adipogenic cocktail (350 nM insulin, 0.5 mM 3-isobutyl-1-methylxanthine, and 250 nM dexamethasone) for 3 d, followed by 3 d in DMEM containing 10% FBS and 350 nM insulin. On day 9 post-differentiation, media volumes were either kept at 1 mL (high) or switched to 0.33 mL (low) for 16 h. Cells were then lysed in 350 µL RLT buffer for RNA extraction and sequencing.