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. 2016 Sep 13;5(9):e362. doi: 10.1038/mtna.2016.67

Figure 7.

Figure 7

Detection of 5′ trans-spliced Dynamin 2 mRNA and protein in muscles. (a) Muscle integrity after AAV-mediated AS1-5′-PTM transduction; Histogram showing the area of muscle sections with internal or central nuclei (%). The area of fibres with internal nuclei was normalized by the total area of the muscle section (Mean + SD, n = 3 sections counted for each condition). (b) RT-PCR analysis on mRNA extracted from transduced muscles using specific primers for trans-spliced and cis-spliced mRNA TS: Trans-spliced PCR amplicon, Endo: endogenous. (c) Electropherogram of trans-spliced amplicons showing Ex13 optimized sequence followed by ex13 bis (isoform 2, top) or ex14 endogenous sequence (isoform 4, bottom). The orange squares show the optimized nucleotides (d). Immunofluorescence analysis on TA transverse sections from mice injected of AAV-AS1-5′-PTM. Fibres were double-stained with anti-FLAG (red) and anti-DNM2 (green) antibodies. Scale bar = 50 µm. AAV, adeno-associated viruses; DNM2, dynamin 2; PTM, pre-trans-splicing molecules; RT-PCR, reverse transcription-polymerase chain reaction; TA, Tibialis anterior.