Skip to main content
. 2012 May 3;287(26):21796–21805. doi: 10.1074/jbc.M111.304626

FIGURE 6.

FIGURE 6.

Effects on 4E-BP1/eIF-4E mediate ERK activation. In A, 8226 or MM1.S cells treated with IGF-1 (200 ng/ml) ± rapamycin or pp242 for 30 min followed by eIF-4E precipitation and subsequent immunoblot of the precipitate for eIF-4E, 4E-BP1, and eIF-4G presence. In B, similar experiment performed in 8226 cells without presence of IGF-1. In C, empty vector (EV) or HA-eIF-4E- stably transfected MM1.S cells (left panel) or 8226 cells (right panel) treated with increasing concentrations of pp242 followed by immunoblot assay for phospho-ERK, total ERK, and HA tag. In D, empty vector or HA-eIF-4E transiently transfected 8226 cells treated with increasing concentrations of pp242 followed by immunoblot assay for phosphorylated and total ERK or phosphorylated and total MEK. In E, empty vector or eIF-4E-transfected cells treated with increasing concentrations of pp242 followed by immunoprecipitation of c-RAF and assay for raf in vitro kinase activity against MEK substrate. Fold increase in kinase activity calculated as described under “Experimental Procedures.”