Fig. 7. N–terminal sequences confer kinase inhibitor sensitivity to the C–terminal phospho-region. HA-tagged eIF4GI fragments were expressed in 293T cells. Cells were metabolically labeled and incubated in 10% serum (A, C and E) or 10% serum + 25 ng/ml rapamycin (B, D and F) for 1 h. Following lysis, immunoprecipitation was conducted sequentially with anti-HA and anti-eIF4GI antisera. Isolated proteins were gel-purified and mapped. The HA–eIF4GI 550–1560 fragment interacts with eIF4E, while HA–eIF4GI 614–1560 does not.