Figure 5 | Scientific Reports

Figure 5

From: Translational enhancement of target endogenous mRNA in mammalian cells using programmable RNA-binding pentatricopeptide repeat proteins

Figure 5

Evaluation of PPR-eIF4G fusion protein in the translational enhancement of c-Myc mRNA. (A) Effect of c-Myc targeting PPR-eIF4G fusion gene (cP64-4G, cP35-4G, or cP2-4G) on c-Myc protein level examined by HTRF assay in HEK293 cells 2 d after transfection. The results of statistical analysis are shown, N = 6, *p < 0.05, **p < 0.01. Error bar indicates the standard deviation. (B) the cell number examined using automated cell counters in HEK293 cells, same as in (A). (C) Target RNA sequence for PPR-eIF4G fusion gene (cP64-4G, pP32-4G, and pP2-4G). (D) c-Myc RNA level determined by RT-PCR using the transfected cells in (A). β-actin was used as an internal control. (E) RIP-PCR was conducted, as shown in Fig. 1E, to analyze the interaction between the PPR-eIF4G fusion protein (cP3-4G, cP10-4G, and pP15-4G) and the targeted c-Myc mRNA in cultured cells. Mock indicates transfection with the empty vector. (F) RIP-PCR analysis of the cross reactivity of p53-targeting pP10-4G and c-Myc-targeting cP35-4G by detection of p53 and c-Myc mRNA.

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