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Table 1.

List of antibodies for immunofluorescent and immunohistochemical staining.

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Fig 1.

Dual IF staining for BRCA1 and CK8/18 and corresponding digital annotation for BRCA1 in a representative core.

A: Composite image where green fluorescent staining if for epithelial CK8/18, red is for BRCA1, and blue is for nuclear DAPI stain. B: Tissue segmentation. C: Nuclear score map. D: Cytoplasmic score map. The attached legend to the right indicates what each color stands for in the tissue segmentation and score maps.

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Fig 2.

IHC staining and corresponding digital annotation for BRCA1 in a representative core.

A: IHC staining utilizes MS110 mouse monoclonal antibody to assess the level of BRCA1 protein expression. B: Tissue segmentation where the epithelial compartment is pink and the stromal compartment is yellow. C: Nuclear segmentation. D: nuclear score map. Definition of the digital image annotation intensity score: blue = 0, yellow = 1, orange = 2 and red = 3. The attached legend to the right indicates what each color stands for in the tissue segmentation and score map.

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Fig 3.

Dual IF staining for BRCA1 and CK8/18 in representative cases of invasive breast cancer and normal breast tissues.

A and B: Normal breast ducts and glands with positive red IF for BRCA1 in myoepithelial cells (arrowheads) and luminal cells (arrows). C: BRCA1 positive ductal carcinoma. D: BRCA1 negative ductal carcinoma. E: BRCA1 positive lobular carcinoma. F: BRCA1 negative lobular carcinoma.

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Table 2.

Distribution of the demographic and tumor-related factors for patients included in this tissue microarray study.

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Fig 4.

Summary of the nuclear and cytoplasmic BRCA1 staining.

A: A plot of pairwise nuclear versus cytoplasmic continuous digital H-score for BRCA1 showing a good correlation between the two locations. B: Percentage of cases that exhibit weak (H-score = <100), Intermediate (H-score > 100 and <200), and strong (H-score >200) cytoplasmic BRCA1 staining. C: Percentage of cases that exhibit weak, Intermediate, and strong nuclear BRCA1 staining.

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Table 3.

Summary of nuclear, cytoplasmic and N/C ratio of BRCA1 expression in normal vs. invasive breast tissue.

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Fig 5.

Dual IF vs IHC staining for BRCA1 analysis.

Plot of pairwise dual IF versus IHC digital H-score for BRCA1 showing a good correlation between the two staining methods for cytoplasmic scores (A), nuclear scores (B), and nuclear/cytoplasmic ratios (C). Figures D to G demonstrate the correlation between the IHC staining (Figs. D and F) and the dual IF staining (Figs. E and G) in the corresponding cores.

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Table 4.

Summary of nuclear, cytoplasmic and N/C ratio of BRCA1 expression among invasive breast cancer samples (IF staining).

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Fig 6.

Association between BRCA1 and other prognostic markers.

A: Positive BRCA1 staining in an invasive ductal carcinoma core. B, C, and D: Positive Her2, CK5/6 and AR staining in corresponding sections of the same core.

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Table 5.

Associations between nuclear, cytoplasmic, and nuclear/cytoplasmic ratio of BRCA1 expression (IF staining) and clinicopathologic features in breast cancer.

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Table 6.

Summary of nuclear, cytoplasmic and N/C ratio of BRCA1 expression among invasive breast cancer samples (IHC staining).

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Table 7.

Associations between nuclear, cytoplasmic, and nuclear/cytoplasmic ratio of BRCA1 expression (IHC staining) and clinicopathologic features in breast cancer.

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