Fig 1: Pharmacological inhibition of SHC1 abrogates OSM-stimulated adipocyte lipolysis. (a) Fully differentiated 3T3-L1 adipocytes were pretreated with the SHC1 inhibitor, PP2 (10 μM), in a regular maintenance medium the night before the assay. On the day of the assay, adipocytes were placed into an incubation medium containing V, the SHC1 inhibitor PP2 (10 μM), 0.5 nM OSM, OSM + PP2, or 10 μM ISO as a positive control, for 4 h. Glycerol measurements were obtained from 50 μL media (n = 7–8 wells per condition). PP2 alone did not affect basal lipolysis, but completely inhibited the OSM induction of lipolysis. (b) Immunoblotting of cell lysates from (a) was used to verify SHC1 inhibition using p66Shc and ERK phosphorylation as readouts. This experiment was conducted twice on independent batches of adipocytes. Significant differences are denoted as follows: **** p < 0.0001 between groups.