Figure 8.

Loss of CypA Reduces Tumor Burden and Lung and Lymph Node Metastases

(A) Representative images of H&E-stained mouse mammary gland containing tumor as a result of cross-breeding the WT (PyMT⁺) mouse to the Cyp A^−/- mouse (n = 12). Different ages are representative of distinct cancer stages in the WT (PyMT⁺) model: 5 weeks, hyperplasia; 8 weeks, adenoma; 11 weeks, invasive carcinoma; 13 weeks, metastasis. Scale bar, 200 μm.

(B) Tumor burden was assessed as the sum of all primary mammary tumor volumes normalized to body weight in grams.

(C) Positivity of lymph nodes was determined as lymph nodes containing at least 1 macrometastasis from FFPE H&E-stained axillary and accessory axillary lymph nodes, assessed by C.V.C., a board-certified pathologist, under bright-field microscopy.

(D) Lung metastases from H&E-stained FFPE lung sections were counted.

(E) Borders were drawn of metastases identified in (D), and a scaled area was calculated for each metastasis as well as each lung lobe for each mouse. Total metastatic area was normalized to total lung area.

(F) Lung metastasis were counted and normalized per tumor burden. All experiments contained n = 12 mice per cohort. For (B and C), data reported as means with SEM. For primary tumor (B) and percent of total positive lymph nodes analysis (C), one-way ANOVA with Tukey's tests for multiple comparisons were used to determine significance.

For (D–F) medians with range were reported due to the lack of a normal distribution of the metastasis data. A non-parametric test for three groups was performed due to the lack of a normal distribution of the data. ∗∗p < 0.01, ∗∗∗p < 0.005, ∗∗∗∗p < 0.001.