Adipogenesis inhibition of 3T3L-1 cells. 3T3L1 cells were maintained in DMEM supplemented with 10% FBS and penicillin-streptomycin. When the cells were confluently grown adipogenesis was initiated by adding IBMX, Dexamethasone, and insulin to 0.5mM, 1 uM, and 10 ug/ml, respectively and continued for 2 days (day 0). The medium was replaced every 2 days with new medium containing insulin in the presence or absence of 5 ug/ml of human Jagged1-Fc and human CD137-Fc as a control-Fc. Staining with Oil Red O was typically performed on day 7.

Adipogenesis inhibition. Macroscopic pictures.

Induction of Hes-1 with the treatment of recombinant human Jagged1-Fc (Prod. No. AG-40A-0081). A mouse preadpipocyte cell line, 3T3L1, was stimulated with 5 ug/ml of human Jagged1-Fc as in indicated time points and each cell lysate was prepared and subjected to western blot by using anti-human Hes1 or GAPDH. Apparently, for Notch ligands or receptors it is normally coated. Otherwise the proteins would need to be artificially multimerized. Since human and mouse Jagged-1 are almost identical (>96%) human works on mouse cells and vice versa.