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. 2008 Oct;14(10):2170–2182. doi: 10.1261/rna.1171808

FIGURE 2.

FIGURE 2.

Effect of eIF4G cleavage and inducible eIF4GI-b and Ct expression on c-myc 5′ UTR-mediated translation. (A, left panel) Uncapped c-myc 5′ UTR-driven reporter translation in untreated (−), CBV3-infected (CBV3), 2Apro co-transfected (2Apro) HeLa cells, or stable tet-inducible eIF4GI-b and Ct expressing cell lines. Cells were infected with CBV3 30 min prior to reporter transfection or reporter RNA was cotransfected with 2Apro expression RNA (Dobrikova et al. 2006). Tet induction was initiated 16 h prior to reporter transfection and all cells were lysed 6 h thereafter. (A, right panel) Capped c-myc 5′ UTR reporter translation in tet-inducible eIF4GI-b and Ct expressing cell lines. (B) Structure of c-myc 5′ UTR reporters. All constructs contained the c-myc 5′ UTR. Gray boxes symbolize a stable stem–loop structure followed by a spacer to block scanning (Thoma et al. 2004; Hundsdoerfer et al. 2005). m7G indicates the presence of a cap structure. (C) Fold stimulation of translation upon tet induction of Ct-expressing cells transfected with the indicated reporter RNAs.