One commercially available TMA slide (HBre-Duc140Sur-01, Shanghai Outdo Biotech Co., Ltd.) was purchased for IHC analysis, which contained histologically confirmed breast cancer tissues with clinico-pathological information, such as tumor grade, clinical stage and the status of ER, PR, and HER2 in IHC (Table 1 ). Breast tumors with positive status of ER or PR belong to luminal-type, and tumors that do not express ER, PR, and HER2 are TNBC. Due to tissue shedding of 15 cases, the number of actually available tissue points was 125. To evaluate the protein abundance of EYA2 in ER– vs. ER+, PR– vs. PR+, and luminal-type vs. TNBC tissues as well as the prognostic value among breast cancer population, IHC analysis was conducted with a standard protocol described previously (23 (link)). The specific primary antibody against EYA2 (ab95875, Abcam) was utilized for IHC at a dilution of 1:100.
Two experienced pathologists performed IHC scoring independently with no prior knowledge of the clinico-pathological information. The multiplication of intensity and proportion of positive-staining tumor cells was exploited to quantify the protein levels of EYA2 according to a standard protocol as described previously (24 (link)).
Two experienced pathologists performed IHC scoring independently with no prior knowledge of the clinico-pathological information. The multiplication of intensity and proportion of positive-staining tumor cells was exploited to quantify the protein levels of EYA2 according to a standard protocol as described previously (24 (link)).
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