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. 2017 Sep 20;7(16):4041–4056. doi: 10.7150/thno.19989

Figure 1.

Figure 1

Notch3 expression is associated with the luminal epithelial phenotype in breast cancer in cell lines and clinical cases. (A) Expression of Notch3, ERα, and EMT-related markers, such as E-cadherin and vimentin, was detected by western blot in breast cancer cell lines MCF-7, T47D, MDA-MB231, SKBR3 and BT549. (B and C) Relative ERα and Notch3 mRNA levels were quantified in breast cancer cell lines by real-time PCR. (D) N1ICD, N2ICD and N4ICD protein levels were detected by western blot in breast cancer cell lines. (E, F and G) Notch1, Notch2 and Notch4 mRNA levels were determined by real-time PCR in breast cancer cell lines. (H) Immunofluorescence staining for Notch3 and ERα was performed in MCF-7 (upper panel) and T47D (bottom panel) cells, and nuclei were stained with DAPI (4', 6-diamidino-2-phenylindole). A merged image was obtained by combining Notch3, ERα and DAPI fluorescent staining images. The scale bar represents 50 μm. (I) ERα and Notch3 protein expression were immunohistochemically stained in ERα-positive (a, b) and ERα-negative (c, d) human breast cancer tissues.