RNA Mis-Splicing Effects of Noncanonical Splicing Variants in Limb-Girdle Muscular Dystrophy Type R1/2A

Figures

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  Figure 1. Transcriptional Analysis of the Noncanonical Splicing Variants c.2185-14T > G and c.1193 + 30G > A (A) DNA electrophoresis presented a 435-bp and a 526-bp band in proband 1 carrying the c.2185-14T > G variant. (B) Sequencing of the upper band (526 bp) revealed retention of the entire intron 20 (91 bp) induced by the c.2185-14T > G variant. (C) DNA electrophoresis presented a 121-bp and a 152-bp band in proband 2 carrying the c.1193 + 30G > A variant. (D) Sequencing of the upper band (152 bp) revealed retention of the first 31 bp of intron 9 caused by the c.1193 + 30G > A variant. M = marker; NC = normal control; P1 = proband 1; P2 = proband 2.

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  Figure 2. Transcriptional Analysis of the Noncanonical Splicing Variants c.1194-9A > G, c.1354 + 5G > A, and c.946-29T > C (A) The pMD18-T cloning and sequencing revealed aberrant spliced transcripts with retention of the last 8 bp of intron 9 induced by the c.1194-9A > G variant. (B) The pMD18-T cloning and sequencing showed aberrant spliced transcripts without the last 40 bp of exon 10 induced by the c.1354 + 5G > A variant. (C) DNA electrophoresis presented a 411-bp band and an 801-bp band in proband 5. (D) Sequencing of the upper band (801 bp) revealed aberrant spliced transcripts with insertion of the last 390 bp of intron 6 induced by the c.946-29T > C variant. M = marker; NC = normal control; P5 = proband 5.

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  Figure 3. Full Western Blots Revealed a Deficiency of 60-kDa Autolytic Fragments of Calpain 3 Protein in Skeletal Muscle Samples From 5 Probands Compared With the Normal Control An additional band of approximately 33 kDa was detected. For proband 3, both calpain 3 and GAPDH protein signals were absent.