Figure 3.

praja2 decreases KSR1 levels. (a) Immunoblot of lysates from cells transiently transfected with Flag-praja2 vector (either wild type or RING mutant). (b) Quantitative analysis (mean±S.E.M) of three independent experiments shown in panel (a) **P<0.01. (c) Cells were transfected with control (siRNAc) or SMARTpool siRNApraja2. Twenty-four hours after transfection, cells were either left untreated or stimulated with Fsk (40 μM, 30 min). To prevent effects of Fsk on protein synthesis, cells were pretreated with cycloheximide. (d) Quantitative analysis of three independent experiments shown in panel (e). *P<0.05 versus control (siRNAc), untreated cells. (e) Immunoblot of lysates from cells transfected with flag-praja2 vector (either wild type or praja2^S342A,T389A). Twenty-four hours after transfection, cells were either left untreated or stimulated with Fsk (40 μM) and harvested at the indicated time points