Fig. 5: Analysis of the MYH3 splicing abnormalities in families D, E, F, and J.

a Schematic representation of the effect of c.1582−6A>G in Family D. b Agarose gel electrophoresis does not show difference in the product size because the length difference of the products is only 5 bp. c The c.1582-6A>G leads to an insertion of 5-bp of intronic sequence before exon 16, leading to a frameshift. d Schematic representation of c.3249-9_3249-5delTCTTC on molecular consequences on cDNA in family E. e Agarose gel electrophoresis illustrating the effect of the splicing variant c.3249-9_3249-5delTCTTC with a larger fragment size in the mutant allele. f Note that the pathogenic variant leads to retention of intron 25, and an in-frame insertion. g Schematic representation of c.5658+3_5658+6del molecular consequences on cDNA in family F. h Sanger sequencing of cDNA from the affected individual and her healthy parents shows that the c.5658+3_5658+6del leads to 9 bp intron retention between exon 39 and exon 40. In silico translation tool in SIB ExPASy Bioinformatics Resources Portal predicts a 9 bp inclusion, adding two amino acids followed by a termination codon annotated by * in the picture. i Schematic representation of c.4357-10G>A molecular consequences on cDNA in family J, showing that the variant leads to exon 32 skipping. j Sanger sequencing of cDNA from the affected individual shows end of exon 31 (single peaks in the chromatogram) and its boundaries to exon 32 and 33 respectively at the start of double peaks in chromatogram.