Fig. 2: Molecular analyses of FBN1. | Human Genome Variation

Fig. 2: Molecular analyses of FBN1.

From: A case of Marfanoid-progeroid-lipodystrophy syndrome: experimental proof of skipping exons and escaping nonsense-mediated decay

Fig. 2: Molecular analyses of FBN1.The alternative text for this image may have been generated using AI.

a Scheme of the genomic variant located in intron 65. b Sanger sequencing of genomic DNA. The heterozygous c.8226+5G>A variant was identified in the proband. c Scheme of PCR in cDNA. PCR primers A and B are indicated by arrows. The expected sizes of the amplicon are 405bp in wild-type and 230 bp in exon 65-skipped mRNA. d Gel electrophoresis of PCR products. PCR was performed on cDNA synthesized from the mRNA of cycloheximide (CHX)-treated and untreated lymphoblastoid cell lines. Extra small bands were observed in the proband, which is consistent with the size skipped by exon 65, as well as large bands common to parents that are considered consistent with the wild-type. e Sequencing analyses of cDNA. cDNA sequences were read in both directions using primers A (forward sequencing) and B (reverse sequencing) with the PCR products. In the proband, two overlapping chromatograms were observed from the junctions of exons 64/65 and 65/66, indicating the skipping of exon 65. The exon-65-skipped mRNA did not seem to be degraded by NMD because CHX treatment did not change the wave heights of the double chromatograms.

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