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. 2009 Sep;175(3):1246–1254. doi: 10.2353/ajpath.2009.090042

Figure 4.

Figure 4

EZH2 overexpression induces intraductal epithelial hyperplasia. A, a: H&E-stained mammary gland tissue sections of virgin MMTV-EZH2 and wild-type (wt) mice at 4 months of age. Mammary specific expression of the MMTV-EZH2 transgene induces a multilayered and disorganized ductal epithelium with formation of epithelial tufts. b: Proliferating cell nuclear antigen (PCNA) immunostaining of 4-month old MMTV-EZH2 and wild-type (wt) mammary glands showing that EZH2 up-regulation increases the proliferation of epithelial cells. Error bars indicate mean ± SD, Student’s t-test, *P < 0.001. B, a: Real-time PCR analysis shows elevated GATA-3 mRNA levels in MMTV-EZH2 mammary glands compared with wild-type mice. 4 m, 4 month; L, lactating; 4 I, involution day 4. Samples were prepared in quadruplicate, and the experiment was repeated three times. Student’s t-test, *P < 0.05, error bars = SD. b: Immunoblot analysis of GATA-3 expression level in MMTV-EZH2 transgenic and wild-type mice mammary tissues. Ponceau S staining is used as the loading control. The number underneath each band indicates the fold change in intensity of the corresponding band relative to the control. c: Immunohistochemical staining for GATA-3 in mature ducts of 4-month-old virgin wild-type and MMTV-EZH2 mice. Note that GATA-3 is up-regulated in the nuclei of epithelial cells of EZH2 transgenic mice compared with those of wild-type mice.