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Fig. S1. Loss of E-cadherin is not tolerated in the mouse mammary gland. (A-D) Immune histochemistry of control (Cdh1F/F;Trp53F/F; A and C) and Wcre;Cdh1F/F (B and D) female mice. Shown are H&E, E-cadherin, cytokeratin (CK) 8 and CK14 stainings. Mammary glands did not show abnormalities in virgin (A and B), or at day 17 of pregnancy (DP17) mice (C and D). Original magnification, (200X).
Fig. S2. Inactivation of E-cadherin and p53 inhibits lactation and nursing. Pups were fostered by their Wcre;Cdh1F/F;Trp53F/+ dam (black bars) or by a lactating wildtype control dam (white bars). Weight was measured and plotted longitudinally. As a control, we determined the average weight of pups from wildtype control females (grey bars). Error bars represent s.e.m. Statistical significance was determined using a two-tailed Student's T-Test. †=mice died after measurements.
Fig. S3. Inactivation of E-cadherin and p53 impairs alveo-lobular development during gestation. Comparative histochemistry showing E-cadherin expression in mammary glands from during pregnant (DP17) Wcre;Cdh1F/F;Trp53F/F mice and control (Cdh1F/F;Trp53F/F) animals. Note the loss of E-cadherin expression in the aberrant lobulo-alveolar structures (arrow) in contrast to typical expression of E-cadherin in the normal ductal and lobular structures (arrow head). Original mouse identification numbers are shown in the right bottom corner and may be cross-referenced with Supplementary Table S1. DP=Day of Pregnancy. Original magnification, (400X).
Fig. S4. Estrogen receptor expression in mammary tumors from the Wcre;Trp53F and Wcre;Cdh1F;Trp53F mouse models. Nuclear expression of estrogen receptor (ER) in mammary tumors. (A) ER expression on a Wcre;Trp53F/F mammary tumor diagnosed as AC + SC/CS. (B) Weak ± and focal (f) ER expression on a Wcre;Cdh1F/+;Trp53F/F mammary tumor diagnosed as SC/CS. (C) Absence of ER expression. on a mILC (solid type) from a Wcre;Cdh1F/F;Trp53F/+ female mouse. D-E: A mixed mILC+SC/CS with focal (f) ER expression. (D) represents the SC/CS component, whereas (E) shows a typical mILC 'scar-type' lesion. (F) ER expression on a mILC from a Wcre;Cdh1F/F;Trp53F/F female mouse. ER expression is indicated by arrows; open arrowheads depict ±(f) ER expression and filled arrowheads point to (pre-existing) ductal structures. Original magnification, (200X).
Fig. S5. Comparative immune histochemistry of metastatic mILC. Expression of cytokeratin (CK) 8 (left panels), CK14 (middle panels) and Vimentin (right panels) by metastatic mILC. (A-C) Metastases from different mILCs to a regional inguinal (A), a draining axillary (B) and a distant caudal (C) lymph node (LN). MD=mammary duct. (D and E) Distant metastases to lung (D) and spleen (E). Note the abundant presence of stromal cells in (C) and (E). B=bronchus. Arrows point to metastatic mILC cells. Scale bars are 100 µm.
Table S1. Comparative histochemistry of mammary tumors from the Wcre;Chd1F;Trp53F mouse model.
Table S2. Tumor spectrum and incidence in the Wcre;Cdh1F;Trp53F mouse model.