LLnL stabilizes the phosphorylation of Shc and MAPK. IL-3–depleted Ba/F₃ cells were treated with or without LLnL as indicated, and stimulated with IL-3 (1 ng/mL) for 0 to 2 hours. (A) Extracts from 10⁷ cells were immunoprecipitated with an anti-Shc antibody and immune complexes were separated by SDS-PAGE (10% gel), transferred, and immunoblotted with RC20 antibody, stripped, and reblotted with anti-Shc antibody as indicated. (B) Whole-cell extracts from the same experiment were also prepared by lysis in 0.5% SDS and 50-μg samples were separated by SDS-PAGE (8% gel), transferred, and immunoblotted with phosphospecific MAPK antibody, stripped, and reblotted with MAPK antibody as indicated. The position of phosphorylated Shc (pShc), Shc, phosphorylated MAPK (pMAPK), and MAPK are indicated.