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. 2014 Nov;88(22):13135–13148. doi: 10.1128/JVI.01883-14

FIG 7.

FIG 7

Dap5 does not interact with MNK and does not affect MNK-dependent PVSRIPO translation competence. (A) HEK293 cells with Dox-inducible expression of Dap5-Flag or eIF4G1(682-1600)-Flag (Ct) (8) were treated with Dox (24 h) and subsequently mock (DMSO) or TPA (200 nM) stimulated (2 h). Flag IP of Dap5/eIF4G1(682-1600) and co-IP of MNK1, eIF4A, and eIF3a are shown. The experiment was repeated 10 times; a representative series is shown. A shift in MNK1 electrophoretic mobility after TPA stimulation likely reflects phosphorylation. (B) HeLa (endogenous) eIF4G1 knockdown/(exogenous) wt eIF4G1 or eIF4G1(ΔMNK) knock-in cells were treated as for Fig. 5. In addition, the cells were treated with siCtrl or siDap5 (72 h) prior to infection. PVSRIPO translation was assayed for each condition alongside relevant controls. The assay was repeated twice, and a representative series is shown.