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. 2020 Sep 8;21(18):6553. doi: 10.3390/ijms21186553

Figure 1.

Figure 1

GGAGAA motif acts as a splicing enhancer. (A) Schematic representation of no exonic splicing enhancer (ESE) control (NEC) and putative ESE (putESE) sequences inserted into pcDNA-Dup vector middle exon used in an ESE-dependent splicing assay [32,33]. Larger uppercase letters illustrate inserted sequence, and bold letters show the putative ESE motif. (B) Agarose gel image shows the RT-PCR analysis of spliced transcripts expressed from NEC and putESE constructs. In parallel, positive controls (SC35) containing three binding sites for SR protein SC35 and negative control (BRCAi11) containing no known regulatory elements derived from BRCA2 intron 11 were used. (C) Inclusion of the middle exon was quantified by capillary electrophoresis, and it is shown as mean ± SD of three independent experiments. Statistically significant difference is shown (*** p < 0.001). (D) Scores from different prediction tools indicated potential splicing enhancing or silencing properties of analyzed sequences. Enhancer character shown as positive, neutral as zero, and silencer as a negative number; 6-mers/7-mers with the highest score of the overlapping sequence are shown.