Immunohistochemical Profiling of Breast Tumors

Tumours from the screening and randomised cohorts were assembled in tissue microarrays using a manual tissue arrayer (MTA-1; BeecherInstruments, Inc., Sun Prairie, WI). The pre-treatment process of deparaffinization, rehydration and epitope retrieval of the 4 μm sections was carried out using the PT Link module (Dako, Glostrup, Denmark). Staining procedure with YAP1 antibody (1:25, Cell Signaling Technology Inc., Danvers, MA, cat#4912) was performed using the Autostainer Plus instrument with the Envision Flex programme (Dako). The epitope used for raising the YAP1 antibody includes amino acid 100 (personal communication, Cell Signaling Technology Europe B.V.) and should therefore detect all to date known isoforms of YAP1 [3 (link)]. YAP1 was scored as overall intensity as either absent, weak, intermediate or strong by a research associate (SL) and a pathologist (GL). Expression of ER, Ki-67, cyclin D1 and amplification of CCND1 (randomised cohort) had been scored previously in both the randomised and screening cohorts [35 (link),45 (link),46 (link)].

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Lehn S., Tobin N.P., Sims A.H., Stål O., Jirström K., Axelson H, & Landberg G. (2014). Decreased expression of Yes-associated protein is associated with outcome in the luminal A breast cancer subgroup and with an impaired tamoxifen response. BMC Cancer, 14, 119.

Publication 2014

MTA-1 PT Link module YAP1 antibody Autostainer Plus instrument

Amino acid Antibody Ccnd1 Cyclin d1 Epitope Isoforms Microarrays Pathologist Rehydration Tissue Weak Yap1

Corresponding Organization :

Other organizations : Lund University, Karolinska University Hospital, Karolinska Institutet, Edinburgh Cancer Research, Linköping University, University of Gothenburg, University of Manchester, Sahlgrenska University Hospital

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Variable analysis

independent variables

Tumor samples from the screening and randomised cohorts

dependent variables

YAP1 expression (scored as overall intensity as either absent, weak, intermediate or strong)
Expression of ER, Ki-67, cyclin D1
Amplification of CCND1 (randomised cohort)

control variables

4 μm sections
Deparaffinization, rehydration and epitope retrieval process using the PT Link module
Staining procedure with YAP1 antibody (1:25, Cell Signaling Technology Inc., Danvers, MA, cat#4912) using the Autostainer Plus instrument with the Envision Flex programme

controls

No positive or negative controls were explicitly mentioned in the provided information.

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