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. 2021 Jul 10;11(7):674. doi: 10.3390/life11070674

Figure 3.

Figure 3

Cterm has no effect on mt-tRNALeu(UUR) steady-state level and aminoacylation. (a) Steady-state levels of mt-tRNALeu(UUR) and mt-tRNAGlu in control (C) and MELAS cybrids, either non-transfected (-) or transfected with empty pcDNA6.2 vector (mock) or Cterm-overexpressing vector (Cterm), were determined by high-resolution northern blot analysis with the indicated specific oligonucleotide probes. Each of the three DIG-labelled oligonucleotides detected a band corresponding to the size expected for mt-tRNALeu(UUR) (78 nt), mt-tRNAGlu (72 nt), and 5S rRNA (120 nt, used as loading control), respectively. The asterisk (*) indicates a non-specific band detected by mt-tRNALeu(UUR) probe. (b) The aminoacylation levels of mt-tRNALeu(UUR) in wild-type cybrids (C) and MELAS cybrids [same as described in (a)] were estimated by acidic high resolution northern blot analysis. DIG-labelled oligos, as in (a), were probed to mt-tRNALeu(UUR) and 5S rRNA. Positions of aminoacylated and deacylated tRNA (80 °C for 15 min at pH 8) are indicated on the left and are in agreement with Chomyn et al. [25].