Fig. 3.

Although stable CHO cell lines overexpressing eIF3i or HAeIF3i were not detected, a CHO cell line expressing HAeIF3c and HEK cell lines individually expressing several eIF3 subunits including an eIF3i over-expressing HEK cell line were successfully generated. A. Cell line screens. The HEK cell lines chosen for further investigation are arrowed. B. A CHO cell line expressing HAeIF3c showed increased growth rate, error bars represent 1 SD from at least 3 separate measurements. Similar data was obtained for a second HAeIF3c clone investigated (Supplementary Fig. 2) and (C), increased cap- and IRES-dependent translation of the luciferase reporter pRMF (error bars represent 1 SD, 3 measurements) and increased expression of eIF3c, eIF3i and c-Myc as determined by western blot (inset). Supplementary Fig. 3 is a repeat of this experiment with a different cell line. D. Western blot of extracts prepared from HEK cells stably expressing the indicated eIF3 subunit (v = vector control) probed for the V5 epitope or the indicated proteins. E. Western blot of V5 PDs from extracts of HEK cells expressing the indicated V5-tagged eIF3 subunits and extracted under either native (N) or denaturing (D) conditions.