Immunohistochemical Analysis of Uqcrh Knockout Mouse Organs
For immunohistochemistry on mouse organs, liver, heart and brain were collected from four Uqcrh−/− and four wild‐type mice (two females and two males per group) at 9 weeks of age at the time of necropsy. The tissues were fixed in formalin for 24 h and embedded in paraffin for histological and immunohistochemical analysis. Immunohistochemistry was performed on 2 µm‐tick sections in an automated immunostainer (Discovery®XT, Roche, Penzberg, Germany). Briefly, sections were incubated with primary antibodies against UQCRC2 (1:1,000; Abcam) and VDAC1/porin (1:2,000; Abcam) with a streptavidin–peroxidase detection reagent. As a negative control, the primary antibody was omitted. Digital images were captured with the NanoZoomer® 2.0HT (Hamamatsu, Japan) digital slide scanner. A scoring system was used to quantify the expression levels of UQCRC2 and VDAC1/Porin: 0 = no staining; 1 = weak staining; 2 = moderate staining; 3 = strong staining. The score for each section was calculated as the mean of 4 high‐power fields (Vidali et al, 2017 (link)).
Other organizations :
Technical University of Munich, Helmholtz Zentrum München, Paracelsus Medical University, German Center for Diabetes Research, Heinrich Heine University Düsseldorf, Deutsches Diabetes-Zentrum e.V., Wellcome Centre for Mitochondrial Research, Newcastle University, Manchester University NHS Foundation Trust, Goethe University Frankfurt, Ludwig-Maximilians-Universität München, Tampere University, Jena University Hospital, German Center for Neurodegenerative Diseases, University of Giessen, German Centre for Cardiovascular Research, University of Manchester
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