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Ab hg fe d c Supplementary Figure 1 Supplementary figure 1: Validation of the ERRα antibody used for immunohistochemical analysis. Control MCF7 breast.

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Presentation on theme: "Ab hg fe d c Supplementary Figure 1 Supplementary figure 1: Validation of the ERRα antibody used for immunohistochemical analysis. Control MCF7 breast."— Presentation transcript:

1 ab hg fe d c Supplementary Figure 1 Supplementary figure 1: Validation of the ERRα antibody used for immunohistochemical analysis. Control MCF7 breast cancer cell line (a), ERRα siRNA treated MCF7 cells (b). Representative selection of immunohistochemical staining pattern of ERRα; negative nucleus (c), strong nucleus (d), negative cytoplasm (e), strong cytoplasm (f), medium cytoplasm and medium nucleus (g), and a mix of medium and negative nuclei (h). To validate the ERRα antibody used for immunohistochemistry MCF7 cells (ATCC, LGC standards, UK) cultured in phenol-red free Optimem media supplemented with 4% FBS (Gibco by Life Technologies, UK) were transfected with control siRNA (Silencer® Negative Control No. 1; AM4611) or ERRα -directed siRNA (Ambion by Life Technologies, UK; s4829 and s4830) using the Amaxa Nucleofector system (Lonza, Swizerland; Program P-020) with the nucleofection reagent kit V. Transfected cells were harvested 72 h post-transfection, formalin fixed, dehydrated and paraffin embedded. Sections of 4µm were prepared for immunohistological staining according to the protocol used for TMAs.

2 ER- Cytoplasmic ERRαNuclear ERRα AB HR (BCS): 1.46 (0.49-4.32) HR (RFS): 1.08 (0.47-2.49) HR (BCS): 2.94 (1.19-7.21) HR (RFS): 1.73 (0.86-3.48) Adjuvant untreated Supplementary Figure 2 Supplementary Figure 2: The prognostic value of ERRα protein expression and subcellular location in adjuvant-untreated ER-negative patients. The prognostic value of ERRα levels evaluated according to hormone receptor status in adjuvant untreated patients (A- B). Breast cancer survival (BCS), Hazard ratio (HR); (95% confidence interval).

3 ER+ Cytoplasmic ERRαNuclear ERRα HR (BCS): 1.46 (0.49-4.32) HR (RFS): 1.27 (0.83-1.93) RFS, HR: 0.76 (0.45-1.29), p=0.31 MFS, HR: 0.81 (0.43-1.51), p=0.50 BCS, HR: 0.93 (0.44-1.98), p=0.85 HR (BCS): 0.98 (0.58-1.68) HR (RFS): 1.18 (0.77-1.79) RFS, HR: 0.86 (0.50-1.47), p=0.58 MFS, HR: 0.93 (0.49-1.75), p=0.82 BCS, HR: 1.32 (0.62-2.80), p=0.48 Adjuvant untreated Tamoxifen treated A C B D Supplementary Figure 3 Supplementary Figure 3: ERRα protein expression is not prognostic in ER-positive breast cancer. The prognostic value of ERRα levels evaluated according to hormone receptor status in adjuvant untreated patients (A, C), and in tamoxifen-treated patients (B, D). Breast cancer survival (BCS), Recurrence-free survival (RFS), Metastasis-free survival (MFS), Hazard ratio (HR); (95% confidence interval).

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