Table 2.

The Immunohistochemical Scoring System Used in Scoring of the Markers' Protein Expression

Marker	Subcellular Localization	Scoring System	Total Score	Criteria	Results
MCM2	Nuclei	% of positively stained nuclei	0%-100%	% of positively stained tumor cells among the total number of malignant cells assessed	MCM2 low = 0%-<35% MCM2 high ≥35%-100%
		IHS	0-12	Combining an estimate of the percentage of immune-reactive cells (quantity score) with an estimate of staining intensity (staining intensity score), by multiplying both scores. Quantity score: 1 (no staining or <10%); 2 (10%-50%); 3 (50%-70%); 4 (70%-100%). Staining intensity score: 0 = negative; 1 = weak; 2 = moderate; 3 = strong.	IHS >3 was considered as positive expression.
		Visiomorph Tissuemorph	0-1	Visiomorph retains only cancer cells in the ROI. Tissuemorph allows accurate counting of the positive and negative nuclei in the ROI.
MCM4	Nuclei	% of positively stained nuclei	0%-100%	% of positively stained tumor cells among the total number of malignant cells assessed	MCM4 low = 0%-<40% MCM4 high ≥40%-100%
		IHS	0-12	Quantity score × staining intensity score	IHS >3 was considered as positive expression.
		Visiomorph Tissuemorph	0-1	Visiomorph retains only cancer cells in the ROI. Tissuemorph allows accurate counting of the positive and negative nuclei in the ROI.
MCM6	Nuclei	% of positively stained nuclei	0%-100%	% of positively stained tumor cells among the total number of malignant cells assessed	MCM6 low = 0%-<55% MCM6 high ≥55%-100%
		IHS	0-12	Quantity score × staining intensity score	IHS >3 was considered as positive expression.
		Visiomorph Tissuemorph	0-1	Visiomorph retains only cancer cells in the ROI. Tissuemorph allows accurate counting of the positive and negative nuclei in the ROI.
Ki-67	Nuclei	% of positively stained nuclei	0%-100%	% of positively stained tumor cells among the total number of malignant cells assessed	Ki-67 low = 0%-<14% Ki-67 high ≥14%-100%
		IHS	0-12	Quantity score × staining intensity score	IHS >3 was considered as positive expression.
		Visiomorph Tissuemorph	0-1	Visiomorph retains only cancer cells in the ROI. Tissuemorph allows accurate counting of the positive and negative nuclei in the ROI.
ER and PR	Nuclei	Allred score	0-8	Sum of the proportion and average intensity scores of positive tumor cells	Negative = 0-2; positive = 3-8
HER-2	Membrane	CAP-approved scoring system	0, 1+, 2+, 3+	0 = no immunostaining or membrane staining which is incomplete or barely perceptible within ≤10% of the invasive tumor cells. 1 + = incomplete membrane or barely perceptible staining within >10% of invasive tumor cells 2 + = circumferential membrane staining that is incomplete and/or weak/moderate within >10% of the invasive tumor cells or complete membranous staining that is intense within ≤10% of the invasive tumor cells. 3 + = circumferential membranous staining that is complete and intense in >10% of tumor cells.	0-1+ are negative; 2+ is equivocal; 3+ is positive