Fig. 7.

In vivo comparison of LICA and the unconjugated parent ASO. We treated TR;HSA^LR mice with LICA oligo 992948 (LICA) or ASO 445236 (ASO), which is the unconjugated parent of LICA oligo 992948 that targets the identical ACTA1 sequence⁸. ASOs were administered by subcutaneous injection of 12.5 mg/kg twice weekly for 4 weeks (N = 3 each) (eight total doses). Treatment with saline (N = 1) served as a control. a DsRed/GFP quantitative fluorescence in gastrocnemius (left) and lumbar paraspinal muscles (right) by serial in vivo spectroscopy in mice treated with saline (black circles), ASO (yellow triangles), or LICA (blue diamonds). Error bars indicate mean ± s.e.m. b Due to the higher baseline DsRed/GFP values in the gastrocnemius of mice randomized to receive the unconjugated ASO, we normalized each measurement of quantitative fluorescence in the gastrocnemius (left) and lumbar paraspinal muscles (right) to the Day 0 value, so that the Day 0 value for each muscle = 1. Error bars indicate mean ± s.e.m. c ddPCR quantitation of ACTA1 transcripts (copies per microliter cDNA) in muscles collected at imaging Day 28. ****P < 0.0001; **P < 0.01; two-way ANOVA. d RT-PCR analysis of exon 22 alternative splicing of the transgene (Tg) and mouse Atp2a1 in gastrocnemius, lumbar paraspinal, quadriceps, and tibialis anterior (TA) muscles collected on imaging Day 28. FVB WT gastrocnemius served as a control. L DNA ladder, bp base pairs. e Quantitation of splicing results in d. Error bars indicate mean ± s.e.m. ****P < 0.0001; **P < 0.01; *P < 0.05; two-way ANOVA