FIGURE 3.
Targeting translation through S6K1. (a) rRNA polysome assembly in PTEN-deficient FL5.12 cells cultured –growth factor for 3 hours ± 4 μM AD80, n=4. (b) 3H-leucine incorporation into the TCA-precipitated protein fraction from PTEN-deficient FL5.12 cells treated ±4 μM AD80. Measurements are mean±SD cpm from triplicate wells cultured in parallel and standardized to Vehicle control. n=5. (c) S6K1−/− MEFs reconstituted with WT-S6K1 or S6K1 T389E were incubated with vehicle control, 20 nM rapamycin or 4 μM AD80. Measurements are as in (b), n=4. (d) Immunoblot analysis of cells cultured as in (c) for 2 hours reveals that S6K1 T389E sustained phosphorylation of the S6K1 substrates rpS6 and eIF4b, indicating a mechanism for AD80 regulation of S6K1-dependent translation control; n=3.