FIG 5.

eIF4E(S209A) substitution and hnRNP A1 depletion do not affect TPA stimulation of PVSRIPO translation. (A) Dox-inducible cell lines expressing wt myc-eIF4E or myc-eIF4E(S209A) were treated with siCtrl (lanes 1 and 5) or eIF4E-targeting siRNA (lanes 2 to 4 and 6 to 8). Dox induction reconstituted wt eIF4E or eIF4E(S209A) to roughly endogenous levels. (B) The assay conditions from panel A, lanes 3 and 4 and lanes 7 and 8, were used to track viral translation after TPA stimulation in the presence of wt myc-eIF4E or myc-eIF4E(S209A) at 3.5 and 4 h p.i. (C) HeLa cells were mock or TPA stimulated following siCtrl or hnRNP A1 siRNA, as shown. The cells were infected, and viral protein was analyzed by immunoblotting (3.5 and 4 h p.i.). The experiments were performed in triplicate (B) or duplicate (C); representative series are shown.