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. 2020 Jul 9;11:3501. doi: 10.1038/s41467-020-17093-9

Fig. 2. ASO walk targeting the NMD-inducing exon inclusion in PCCA.

Fig. 2

a Diagram of ASO walk. The target region spans 104 nt of intronic sequences on each side of the non-productive alternative exon 14X, the exon–intron boundaries and exon 14X. Black lines denote introns, gray rectangles denote constitutive exons, and red rectangle denotes non-productive alternative exon. b A representative RT-PCR PAGE from HEK293 cells transfected with a total of 54 18-mer ASOs (shown in panel a) and a non-targeting ASO control (NT) at 80 nM concentration or no-ASO mock control (−) for 24 h. The top band (195 bp) corresponds to the non-productive mRNA that results from the inclusion of exon 14X and the bottom band (111 bp) corresponds to the productive mRNA that results from the skipping of exon 14X (n = 2 biologically independent samples over two independent experiments). c Graph depicting the fold increase over no-ASO mock control (−) of PCCA productive mRNA determined by TaqMan qPCR (gray circles, n = 2) and the fold decrease of PCCA non-productive mRNA based on densitometric quantification of RT-PCR products (red squares, n = 2) from panel b. Data are presented as mean values ±SD. Results show an inverse correlation between the reduction of non-productive mRNA and the increase of productive mRNA. ASO sequences are in Supplementary Data 4. Source data are provided as a Source Data file.