Figure 5.

Proteins synthesized during heat shock mediate insolubilization of eIF4G. (A) 293T cells were treated with cycloheximide at 50 μg/ml to prevent protein synthesis, then lysed in nonionic (Triton X-100) buffer to solubilize proteins at the indicated times. (B) 293T cells transfected with GST–Mnk1 were treated with 50 μg/ml cycloheximide as shown, lysed in Triton X-100 buffer, and GST–Mnk1 was purified by glutathione–Sepharose chromatography. Proteins were resolved by SDS-PAGE and immunoblotted with specific antisera as shown.