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. 2017 Oct 3;46(1):e1. doi: 10.1093/nar/gkx867

Figure 6.

Figure 6.

Responsivity of cDNA slot blot and alternative splicing to treatment with antisense oligonucleotides (ASOs) in transgenic mice. (A) cDNA slot blot of 2 μg total RNA from quadriceps muscle (HSALR and HSAXLR mice) or left ventricle (LC15 mice) treated with subcutaneous injections of saline (n = 4) or ASO (n = 4 in each group). Negative control samples from HSASR quadriceps, HSANR quadriceps, or wild-type (WT) heart are indicated. (B) Relative ER-RNA quantification by cDNA slot blot (black bars) versus qRT-PCR (white bars). Saline-treated groups set to 1 for both assays. (C) RT-PCR analysis of alternative splicing in saline- versus ASO-treated HSALR quadriceps (Serca1 and Clcn1) and LC15 heart tissue (Tmem63b and Cacna1s), as compared to wild-type controls. The mean ± S.D. values of ‘percent spliced in’ for saline versus ASO versus wild-type groups were Serca1 exon 22: 15 ± 2%, 94 ± 2%, 100 ± 0%; Clcn1 exon 7A: 31 ± 2%, 5 ± 1%, 4 ± 2%); Tmem63b exon 5: 46 ± 4%, 66 ± 2%, 84 ± 0%; Cacna1s exon 29: 38 ± 1%, 78 ± 1%, 95 ± 1%). (D) Quantification of (C). ANOVA of Serca1 exon 22 (P = 2.2 × 10−9), Clcn1 exon 7A (P = 4.4 × 10−6), Tmem63b exon 5 (P = 3.1 × 10−9), and Cacna1s (P = 3.7 × 10−9). *Student's t-test versus WT P < 0.0005, † = Student's t-test versus ASO P < 0.00005.