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01-May-2016 05:34

Presence of S105-ERbeta in breast cancer and association with improved survival, even in endocrine resistant breast tumors suggest S105-ERbeta might be a useful additional prognostic marker in this disease.Although ERα phosphorylation correlates with patient outcome, ERβ is similarly phosphorylated but its significance in breast cancer has not been addressed.Total ERβ (14C8, Ab Cam; 2.5 μl) and P-S105-ERβ (Ab Cam; 5 μl) were added to the cell suspension and incubated for 30 minutes at 4°C.Labeled cells were washed twice and incubated for 1 hour at 4°C with fluorescein isothiocyanate-conjugated secondary antibodies (rabbit anti-mouse , swine anti-rabbit 0; both Sigma). Samples were stored overnight in the dark at 4°C then analyzed using an LSRII flow cytometer (BD, Oxford, UK) on pre-stored instrument settings.

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Nuclear S105-ERbeta was observed in breast carcinoma and was associated with better survival (Allred score or =3), even in tamoxifen-resistant cases, and additionally correlated with ERbeta1 and ERbeta2 expression.Blots were incubated at 4°C for 15 hours with S105-ERβ antibody, ERβ1 (Serotec, Oxford, UK; both 0) or 00 S118-ERα (Cell Signaling, Danvers, MA), all diluted in 0.5% bovine serum albumin.

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