Figure 2.

Translation efficiency and accuracy of multiple modified mRNA nucleotides. (A) Sequence of the reporter eGFP mRNA is depicted. Employing in vitro transcription, the Us and Cs were substituted with Ψs and m⁵Cs, respectively. The respective Cs are depicted in green and the Us in blue. The amino acids corresponding to selected codons (red boxes) were analyzed by mass spectrometry for their identity (detection limit ~1%). (B) Western blot analysis of the translation products derived from mRNAs carrying multiple Ψs or m⁵Cs. (C) Western blot analysis of translation products harboring one, two or three m⁶As.