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Fig. 2 | Skeletal Muscle

Fig. 2

From: TRIM32 biallelic defects cause limb-girdle muscular dystrophy R8: identification of two novel mutations and investigation of genotype–phenotype correlation

Fig. 2

Molecular genotyping and expression analysis of mutated TRIM32 at the RNA and protein levels in vivo and in vitro. A Detection of the deletion in TRIM32 by multiplex gap-PCR and the exact deletion breakpoint of TRIM32 in the proband (chr9.hg19:g.119431290_119474250del). B Sanger sequencing of TRIM32 in the proband (c.1700A > G). C Multiplex sequence alignment in various species revealed that the variable base (567, labeled in red) was relatively conserved. D The predicted 3D models of the wild-type (right) and mutated (left) TRIM32 protein. An area different between the two was marked after contrastive analysis by PyMOL. E HEK 293 T cells transfected with a wild-type plasmid (pEGFP-TRIM32-WT) or mutant plasmids (pEGFP-TRIM32-D487N; pEGFP-TRIM32-H567R) under the same conditions showed that the mutant constructs exhibited diffuse cytoplasmic staining with loss of characteristic aggregates. Scale bar = 100 µm (× 40 magnification). F Western blot analysis of TRIM32 protein showed that the upper band corresponding to TRIM32/ubiquitin appeared in the wild-type and not in the mutant types (p.D487N and p.H567R). G Quantitation of TRIM32 gene transcription levels in the family

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Skeletal Muscle

ISSN: 2044-5040

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