Figure 7.

The overexpressed EV-A71 3D elevates IRES-driven translation in vivo. (A) Scheme shows the bicistronic RNA produced from pRHF-EV-A71 reporter plasmid. Translation of firefly and Renilla luciferase are dependent on 5ʹ end cap and EV-A71 IRES, respectively. (B) Reporter plasmid was co-transfected with effector plasmid encoding EV-A71 3D proteins into HEK293T cells. The overexpressed 3D of EV-A71 C2 (lane2) and C4 (lane3) strain enhanced the EV-A71 IRES activity. Expression level of 3D proteins was examined by immunoblotting using anti-FLAG antibody. β-actin served as the loading control. (C) Similar to the experiment described in (B) but control siRNA (Ctrl) or siRPS6 was included. Knockdown of RPS6 was confirmed by immunoblotting with anti-RPS6 antibody. The error bars represented the standard deviation (SD). Statistical analysis was performed by the Student’s t test. ***: p <0.001; **: p <0.01. ns: not significant.