Journal: Frontiers in Genetics

Article Title: Case Report: Incidental late-onset Pompe disease diagnosis in a man with no clinical and instrumental evidence of neuromuscular dysfunction

doi: 10.3389/fgene.2025.1574381

Figure Lengend Snippet: Genetic studies (A) Pedigree of the family investigated. Subjects harbouring IVS1-32-13T>G in homozygosis (+/+) or heterozygosis (+/−) are shown. The arrow indicates the Proband. (B) Reverse transcription PCR (RT-PCR) encompassing GAA transcript Exons 1 (E1) and 3 (E3) in lymphocytes of the subjects indicated. Amplicons corresponding to normally spliced molecules including Exon 2 (“E2” = Normal transcript “N”) and altered transcripts (Splice Variants “SV1-3”) are indicated. (C) Tape Station analysis of RT-PCR amplicons and their relative quantification. (D) Quantitative RT-PCR analysis of GAA transcripts in lymphocytes of the subjects indicated as evaluated by Taqman probes targeting Exons 1-2 and 4-5 junctions.

Article Snippet: The following probes were used: Hs00164635_m1 ( GAA , exon junction 1-2); Hs01089838_m1 ( GAA , exon junction 4-5); Hs99999903_m1 ( ACTB, encoding the housekeeping beta actin).

Techniques: Reverse Transcription, Reverse Transcription Polymerase Chain Reaction, Quantitative Proteomics, Quantitative RT-PCR

Journal: Frontiers in Genetics

Article Title: Case Report: Incidental late-onset Pompe disease diagnosis in a man with no clinical and instrumental evidence of neuromuscular dysfunction

doi: 10.3389/fgene.2025.1574381

Figure Lengend Snippet: Transcript and Protein analysis of Patients’ fibroblasts. (A) Reverse transcription PCR (RT-PCR) encompassing GAA transcript Exons 1 (E1) and 3 (E3) in Proband’s and control fibroblasts. Amplicons corresponding to normally spliced molecules including Exon 2, (“E2” = Normal transcript “N”) and altered transcripts (Splice Variants “SV1-3”) are indicated. (B) Tape Station analysis of RT-PCR amplicons and their relative quantification. (C) Quantitative RT-PCR analysis of GAA transcripts in Proband’s and control fibroblasts as evaluated by Taqman probes targeting Exons 1-2 and 4-5 junctions. (D) Western blot analysis of GAA protein levels in Proband’s and control fibroblasts. Mature forms of GAA enzyme correspond to bands at 76 and 70 kDa. GAPDH levels are used for normalization purpose.

Article Snippet: The following probes were used: Hs00164635_m1 ( GAA , exon junction 1-2); Hs01089838_m1 ( GAA , exon junction 4-5); Hs99999903_m1 ( ACTB, encoding the housekeeping beta actin).

Techniques: Reverse Transcription, Reverse Transcription Polymerase Chain Reaction, Control, Quantitative Proteomics, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Genetics

Article Title: Case Report: Incidental late-onset Pompe disease diagnosis in a man with no clinical and instrumental evidence of neuromuscular dysfunction

doi: 10.3389/fgene.2025.1574381

Figure Lengend Snippet: Genetic studies (A) Pedigree of the family investigated. Subjects harbouring IVS1-32-13T>G in homozygosis (+/+) or heterozygosis (+/−) are shown. The arrow indicates the Proband. (B) Reverse transcription PCR (RT-PCR) encompassing GAA transcript Exons 1 (E1) and 3 (E3) in lymphocytes of the subjects indicated. Amplicons corresponding to normally spliced molecules including Exon 2 (“E2” = Normal transcript “N”) and altered transcripts (Splice Variants “SV1-3”) are indicated. (C) Tape Station analysis of RT-PCR amplicons and their relative quantification. (D) Quantitative RT-PCR analysis of GAA transcripts in lymphocytes of the subjects indicated as evaluated by Taqman probes targeting Exons 1-2 and 4-5 junctions.

Article Snippet: The following probes were used: Hs00164635_m1 ( GAA , exon junction 1-2); Hs01089838_m1 ( GAA , exon junction 4-5); Hs99999903_m1 ( ACTB, encoding the housekeeping beta actin).

Techniques: Reverse Transcription, Reverse Transcription Polymerase Chain Reaction, Quantitative Proteomics, Quantitative RT-PCR

Journal: Frontiers in Genetics

Article Title: Case Report: Incidental late-onset Pompe disease diagnosis in a man with no clinical and instrumental evidence of neuromuscular dysfunction

doi: 10.3389/fgene.2025.1574381

Figure Lengend Snippet: Transcript and Protein analysis of Patients’ fibroblasts. (A) Reverse transcription PCR (RT-PCR) encompassing GAA transcript Exons 1 (E1) and 3 (E3) in Proband’s and control fibroblasts. Amplicons corresponding to normally spliced molecules including Exon 2, (“E2” = Normal transcript “N”) and altered transcripts (Splice Variants “SV1-3”) are indicated. (B) Tape Station analysis of RT-PCR amplicons and their relative quantification. (C) Quantitative RT-PCR analysis of GAA transcripts in Proband’s and control fibroblasts as evaluated by Taqman probes targeting Exons 1-2 and 4-5 junctions. (D) Western blot analysis of GAA protein levels in Proband’s and control fibroblasts. Mature forms of GAA enzyme correspond to bands at 76 and 70 kDa. GAPDH levels are used for normalization purpose.

Article Snippet: The following probes were used: Hs00164635_m1 ( GAA , exon junction 1-2); Hs01089838_m1 ( GAA , exon junction 4-5); Hs99999903_m1 ( ACTB, encoding the housekeeping beta actin).

Techniques: Reverse Transcription, Reverse Transcription Polymerase Chain Reaction, Control, Quantitative Proteomics, Quantitative RT-PCR, Western Blot