Immunohistochemistry and AQUA analyses were performed on sections containing either xenotransplant tissues or a tissue array constructed by cutting edge matrix assembly containing 140 deidentified breast carcinoma specimens (DCIS, primary invasive ductal carcinomas (Grades 1–3), and lymph node metastases) and 40 normal breast tissues (Supplementary Data ). Immunohistochemistry was performed as described previously (44 (link)) using Stat5a (Advantex, 1:8,000), Stat5b (Advantex, 1:4,000), pY-Stat5 (Epitomics, 1:200), and BCL6 (Santa Cruz, 1:50). AQUA analysis was performed using AQUA/PM2000 (HistoRx) (45 (link)). Briefly, slides were scanned and fluorescent images were captured in three channels (FITC/Alexa-488, Cy5, or DAPI). AQUA scores for Stat5a, Stat5b, pY-Stat5 and BCL6 represent average signal intensities within the epithelial cell compartment as defined by cytokeratin-positive mapping.
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Quantifying Protein Biomarkers in Breast Cancer
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Bcl6
Breast
Breast carcinoma
Cytokeratin
Dapi
Dcis
Fitc
Immunohistochemistry
Invasive ductal carcinomas
Lymph node metastases
Normal tissues
Signal cell
Stat5
Stat5b
Corresponding Organization : Sidney Kimmel Cancer Center
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Variable analysis
independent variables
- Tissue type (xenotransplant tissues or tissue array containing DCIS, primary invasive ductal carcinomas (Grades 1–3), and lymph node metastases, and normal breast tissues)
- Stat5a expression
- Stat5b expression
- PY-Stat5 expression
- BCL6 expression
- Slide preparation and AQUA analysis methodology as described in previous work (references 44 and 45)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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