PMC:5985359 / 33324-35192 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/5985359","sourcedb":"PMC","sourceid":"5985359","source_url":"https://www.ncbi.nlm.nih.gov/pmc/5985359","text":"We investigated the potential of (CAG)7 ASO treatment to reverse the shift in alternative splicing events previously observed in expansion-positive CEC lines (Figure 4). Ten FECD expansion-positive CEC lines (Table S9) were treated with either the (CAG)7 or control ASO. Following treatment, RNA was extracted and RT-PCR was performed. For both MBNL1 and MBNL2 there was a highly significant (p \u003c 0.0001) shift in the relative proportions of alternatively spliced transcripts after (CAG)7 ASO treatment (n = 10) toward the control CEC spliceoform distribution (Figure 6). Furthermore, a significant (p ≤ 0.05) shift in the relative proportions of alternatively spliced transcripts was also demonstrated for NUMA1 (n = 10) (Figure 6). These data therefore demonstrate that the (CAG)7 ASO is effective at rescuing the differential splicing events associated with CTG18.1 expansion-related pathology (Figures 4 and 6).\nFigure 6 ASO Treatment Rescues Differential Splicing Events Underlying CTG18.1-Associated Pathology\n(A) Representative RT-PCR images for non-treated samples with either expanded (E) or control (C) (non-expanded CTG18.1 genotypes) are shown on the far left, for reference purposes. Representative RT-PCRs are shown for three independent ASO-treated CEC lines.\n(B) Schematic representations are provided for each respective reaction. Primer locations are denoted with arrows. The respective sizes of all amplified products are given.\n(C) Percentage expression of amplicons of interest (A or B) relative to total amplified products, per reaction, are presented as a mean for control-ASO versus (CAG)7 ASO-treated groups. The mean from 10 independent CEC lines ± SD. In all cases CECs were treated with 200 nM (CAG)7 or control ASO for 24 hr. p values were calculated using a paired two tailed t test for each event investigated; ∗∗∗∗p \u003c 0.0001, ∗p \u003c 0.05.","divisions":[{"label":"label","span":{"begin":916,"end":924}},{"label":"p","span":{"begin":925,"end":1015}},{"label":"p","span":{"begin":1016,"end":1274}},{"label":"p","span":{"begin":1275,"end":1447}}],"tracks":[]}