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222 files

Quadruple immunofluoresence images of skeletal muscle biopsies from patients with the mtDNA m.3243A>G variant

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posted on 2022-04-29, 07:19 authored by Syeda Tasnim Ahmed, Doug Turnbull, Robert Taylor, Conor LawlessConor Lawless, Sarah PickettSarah Pickett

Fluorescent images of 10µm transverse muscle sections from patients carrying the pathogenic m.3243A>G mtDNA variant, captured by automated scanning at 20× magnification using Zen 2011 (blue edition) software and Zeiss Axio imager MI microscope. Exposure times across all sections for each experimental batch were maintained.


Muscle sections were stained using a 'Quantitative quadruple immunohistochemistry' technique, designed to capture expression profiles of proteins belonging to oxidative phosphorylation (OXPHOS) complexes. 'OXPHOS' sections were labelled with antibodies detecting subunits of mitochondrial complex I (NDUFB8) and complex IV (COX-1), a mitochondrial mass marker (mitochondrial porin; VDAC1) and laminin, followed by incubation with secondary antibodies (Alexa Fluor 488, 546, biotinylated IgG1 and 750) and subsequently with streptavidin 647. Alongside each sample, a no-primary control section (NPC; labelled only for laminin) was processed.


This resource contains .iaf, .jpg and .png files from OXPHOS and NPC sections from 17 patients (P01-P17) and five controls (C01-C05). It also contains PDFs for 10 sections for which single fibre molecular genetic data are available at https://github.com/CnrLwlss/Ahmed_2022


File naming convention:


b : where n=2 or 3 and refers to the batch that the samples were processed in. 

P01-P17 : Sections from m.3243A>G patients

C01-C05 : Non-disease controls

NPC : Non-primary control - sections labelled only with laminin

OXPHOS : Sections labelled for OXPHOS complexes

ch1 : MTCO1 (Complex IV; 488nm)

ch2 : VDAC (Porin; 546nm)

ch3 : NDUFB8 (Complex I; 647nm)

ch4 : Laminin (750nm)






Funding

Wellcome Centre for Mitochondrial Research

Wellcome Trust

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Identification of nuclear factors modulating the clinical phenotype of m.3243A>G-related mitochondrial disease.

Wellcome Trust

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MRC Strategic Award to establish an International Centre for Genomic Medicine in Neuromuscular Diseases

Medical Research Council

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History

UoA

  • Clinical Medicine