Data and metadata supporting the published article: S-nitrosylated and non-nitrosylated COX2 have differential expression and distinct subcellular localization in normal and breast cancer tissue

Immunohistochemical staining in breast cancer shows both gain and loss of COX2 expression with disease risk and progression. In this study, the authors investigated the effect of antibody clone selection on COX1 and COX2 recognition, focusing on four commonly utilized αCOX2 clones. They validated three clones, SP21, CX229, and CX294 as highly specific for COX2 protein.
Data access: The data generated and/or analysed during the current study, are publicly available in the figshare repository, as part of this data record (https://doi.org/10.6084/m9.figshare.13009985). The clinical data that support the findings of this study, are available from the corresponding author, Dr. Pepper Schedin (email address: [e-mail]), upon reasonable request.
Study approval and patient consent: Formalin-fixed paraffin-embedded (FFPE) human breast and colon tissue for this study was approved by the BWH/Harvard Cohorts Biorepository and Institutional Review Boards at Colorado Multiple Institution Review Board (COMIRB), and Oregon Health and Science University (OHSU). Written informed consent was given by participants when required.
Study aims and methodology: The cyclooxygenase enzyme COX2, a key mediator of tissue inflammation via prostaglandin production, has been investigated extensively as a cancer biomarker and therapeutic target. In this study, the authors investigated the effect of antibody clone selection on COX1 and COX2 recognition, focusing on four commonly utilized αCOX2 clones.De-identified FFPE cases of breast (n=52) and colon (n=53) cancer tissue microarrays (TMAs) and breast cancer cases (n=2019) from the Nurses’ Health Study-1 (NHS1) were obtained from the Channing Laboratory, Brigham and Women’s Hospital, Massachusetts. Young women’s FFPE breast cancer cases were acquired from the Young Women’s Breast Cancer Translational Program (YWBCTP) at the University of Colorado (n = 233). Breast tissue sections with adjacent normal, ductal carcinoma in situ (DCIS) and invasive ductal carcinoma on a single slide were obtained from Kaiser Permanente Northwest (KPNW) (n=10). A total of 233 YWBCTP and 1770 NHS1 cases were evaluated for dual COX2 IHC stain after exclusion of one entire TMA slide (249 cases) from the NHS1 cohort.The following procedures are described in more detail in the related article: immunoblotting, S-nitrosylation and chemical de-nitrosylation, immunohistochemical staining of FFPE tissues, hierarchical and K means clustering of SP21 and CX229 expression, and statistical analysis.
Data supporting the published article: A list of all the datasets and their file formats is provided in the file Jindal, S. et al.xlsx. The datasets listed are all part of this figshare data record.
Software needed to access data: The .svs files can be opened using the Inkscape software. The .czi files can be opened using Image J or the Carl Zeiss Zen software. The aforementioned software programmes are available as free download.