Tissue Microarray Analysis of Breast Cancer Markers

Tissue microarray (TMA) was collaborated with Alenabio Technology Co., Ltd. (Xian, China). The tissue microarray contained 138 breast cancer specimens. Briefly, paraffin sections were first deparaffinized, antigen retrieval was performed in citrate buffer (pH 6.0), and endogenous peroxidase activity was blocked in 0.3% H₂O₂. The slides were continuously incubated with the indicated primary and secondary antibodies until visualization with peroxidase and 3,3′-diaminobenzidine tetrahydrochloride. The expression of CPEB1, NOTH3, NUAK1, and PDPK1 in the breast cancer tissues from the tissue microarray was blindly quantified by two pathologists based on histochemical score (H-score) as previously described (22 (link)). The primary antibodies were CPEB1 (Proteintech, Wuhan, China 13274-1-AP), NOTCH3 (Proteintech, Wuhan, China 55114-1-AP), NUAK1 (Proteintech, Wuhan, China 22723-1-AP), and PDPK1 (Proteintech, Wuhan, China 17086-1-AP).