Differentiated 3T3-L1 cells were treated with increasing concentrations of isoproterenol for 48 hours and the culture supernatant was assayed for free glycerol. Top panel: glycerol release from undifferentiated (red) and differentiated (blue) 3T3-L1 cells as determined using Free Glycerol Assay Reagent. Bottom panel: isoproterenol dose response curve showing glycerol released after 24 hours versus isoproterenol concentration.
Example of standard curve of typical data produced with 3T3-L1 culture medium.
Upon induction, 3T3-L1 cells differentiate into adipocytes-like cells and they are ready to be used for adipolysis experiments. Panel A: non-differentiated 3T3-L1 cells show no staining of Oil Red O Solution. Panel B: increased lipid droplet accumulation indicates that more than 80% of preadipocytes were differentiated into adipocytes four days after weaning the cells from Induction Medium to Insulin Medium.