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. Author manuscript; available in PMC: 2017 Dec 13.
Published in final edited form as: Cell Rep. 2017 Nov 21;21(8):2039–2047. doi: 10.1016/j.celrep.2017.10.096

Fig. 3. 5’UTRs of Drosomycin, Attacin A and 4E-BP can be translated cap-independently.

Fig. 3

3A. Schematic showing the bicistronic construct (top). (Bottom) Luciferase activity from S2 cells expressing bicistronic constructs containing the 5’UTRs from Drosomycin (Drom), Attacin A (AttA) and 4e-bp, with the empty vector (EV) without any 5’UTR inserted (EV) as a negative control and, Hepatitis C virus (HCV) IRES inserted as a positive control. The mini-Actin 5C promoter (Act5CP) drives reporter expression. Firefly luciferase activity of each sample is normalized to its respective Renilla luciferase activity.

3B. 5’-capped Firefly monocistronic reporter mRNA (schematic on top) translated in control RRL (white) or RRL treated with excess cap-complex (black). Data for each 5’UTR are normalized to respective control samples.

3C. Monocistronic assay as performed in 3B but in RRL treated with microccocal nuclease to eliminate competing capped transcripts.

3D. Comparison of translation efficiency of capped versus uncapped monocistronic mRNA in microccocal nuclease-treated RRL (as performed in 3B).

Error bars indicate standard error from at least 3 independent experiments. **=p<.01. Fold changes are not significant unless indicated otherwise.