Multiway ANOVA (anovan; MATLAB) was performed to examine the relationship between EV density and the corresponding grade of IDC/DCIS. Noticing that the tumor-to-margin distance also contributes to the EV density, we only included in the multiway ANOVA the cases with a tumor-to-margin distance between 0 and 8 mm to minimize the contribution by distance and maintain a sufficient number of data points. Statistically significant correlation was found between the EV density and the histological grade of IDC (P = 0.0002), but not with the nuclear grade of DCIS (P = 0.1835). Furthermore, with statistical significance existing between the EV density and the histologic grade of IDC, a multiple comparison test (multcompare, critical value: “tukey-kramer,” MATLAB) was used to analyze the differences of EV densities between different histologic grades of IDC (Fig. 3E).

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