Adipogenesis inhibition of 3T3L1 cells: 3T3L1 cells were maintained in DMEM, w/10% fetal bovine serum, penicilin-streptomycin. Adipogenesis was initiated by adding 1uM Dexamethasone, 0.5mM IBMX, 10ug/ml lnsulin (day 0) until day 2. The medium was replaced every 2 days w/new medium containing insulin in the presence or absence of 044747 (5ug/ml) and hCD137-Fc (Fc control). Staining w/Oil Red O was performed on day 7. Cells were washed twice w/PBS, fixed w/3.7% formalin, and stained w/0.5% filtered Oil Red O in propylene glycol.
Induction of Hes-1 with the treatment of 044747: A mouse preadpipocyte cell line, 3T3L1, was stimulated with 044747 (5mg/ml) as in indicated time points and each cell lysate was prepared and subjected to western blot by using anti-mouse Hes1 or GAPDH. Lane 1: 0min, Lane 2: 10min, Lane 3: 30min, Lane 4: 1hr, Lane 5: 2hr, Lane 6: 4hr, Lane 7: 8hr, Lane 8: 24hr.
Adipogenesis inhibition. Macroscopic pictures.
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