Figure 8.

Validation of a Differentially Used Junction in COL25A1

(A) Sashimi plot (software Integrative Genomics Viewer) showing the raw reads data mapped on COL25A1 (exons 20–22) in human primary myotubes non-transduced (NT) or transduced with rAAV3b-U7-E53 (U7). Alignments on exons were represented as read densities (spikes), and alignments on junctions were represented as arcs between exons. The exact number of reads mapped on each junction was specified. (B) Splicing modifications of COL25A1 messenger analyses using nested RT-PCR. The COL25A1 messenger was detected from exons 19–23 in three replicates of primary myotubes sequenced through RNA-seq and four supplemental primary myotube samples transduced with rAAV3b-U7-E53 or rAAV3b-U7-Scr. Two PCR products of 210 and 165 bp were detected, corresponding, respectively, to COL25A1 isoforms with and without exon 21. H2O, water; L, DNA ladder.