Anti-DICER1 antibody produced in rabbit

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The Outcome tissue microarray (TMA) was obtained from the Cooperative Prostate Cancer Tissue Resource (CPCTR) study [27]. This TMA contains cores from radical prostatectomy specimens; 4 cores per patient and each patient with PSA recurrence was matched (by age, grade and stage) to a patient without recurrence [27]. Slides 1 and 2 of the array were used and contained 170 total patients, 85 matched pairs. Five micron sections were deparaffinized, subjected to antigen retrieval, stained with anti-DICER1 1:100 overnight at 4°C (Prestige antibody HPA000694, Sigma-Aldrich, Hamburg, Germany) and visualized with mouse DAB Avidin/Biotin Complex Kit (VectorLabs, Peterborough, UK). Analysis. Areas of epithelium and stroma were marked manually by a pathologist, then segmented and quantified with the Vectra Tissue Imaging System (PerkinElmer, Waltham, MA). Cores that were incomplete, damaged or did not contain cancer epithelium and stroma were excluded. Areas of inflammation were not included. Using the Vectra system, the total DAB intensities for the epithelium and the stroma was determined from the area and pixel intensity. To control for core-to-core staining differences, the epithelial/stromal ratio per core was determined and analyzed by conditional logistic regression using the outcome of biochemical PSA recurrence (BCR). A total of 170 patients were analyzed; N=85 non-recurrent, N=85 for recurrent disease (BCR).

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