IHC was carried out as previously described (n=3 untreated, n=5 JQ1 treated) 35 (link). Slides were submitted to antigen retrieval, and the primary antibodies listed in Table S7 . Slides were incubated with secondary antibody (Dako) and DAB (Dako) and counterstained with haematoxylin solution (Sigma-Aldrich). Secondary-only control staining was done routinely. ImageJ colour deconvolution was used for quantification 35 (link).
Protocol detail
Immunohistochemical Analysis of JQ1 Treatment
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Corresponding Organization : University of Nottingham
Other organizations : MRC Weatherall Institute of Molecular Medicine, University of Oxford, Universidade Federal do Rio Grande do Sul, Wellcome Centre for Human Genetics, University of Plymouth, Genomics England
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Variable analysis
independent variables
- Treatment (untreated vs. JQ1)
- Antigen expression (quantified using ImageJ color deconvolution)
- Antigen retrieval process
- Primary antibodies used (as listed in Table S7)
- Secondary antibody (Dako)
- DAB (Dako)
- Haematoxylin counterstain (Sigma-Aldrich)
- Secondary-only control staining
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