Figure 1 | Scientific Reports

Figure 1

From: Trastuzumab resistance induces EMT to transform HER2+ PTEN− to a triple negative breast cancer that requires unique treatment options

Figure 1

Continued use of trastuzumab in HER2+ breast cancer with PTEN inactivation generates resistance, induces a mesenchymal phenotype and increases cancer stem cells.

(A) Representative bright field microscopy images of BT474, BT474 shPTEN and BT474 shPTEN with long term treatment of trastuzumab (LTT). Original images obtained with 20× objective. (B) Left, percent of cells expressing basal breast cancer markers CD44+/CD24- in cell lines as assessed by flow cytometry. N = 3. Right, representative CD44 and CD24 flow cytometry analysis of each. (C) Real time PCR quantification of epithelial cell makers E-cadherin and EpCAM normalized to expression of GAPDH. (D) Real time PCR analysis of mesenchymal cell markers N-cadherin and Vimentin, expressed relative to GAPDH. N = 4. (E) Western blot analysis of protein level expression of EMT related genes. (F) Mammosphere formation rates of parental BT474 and BT474 PTEN− LTT cell lines. N = 3. (G) Representative image of macrometastasis that forms in 1 in 5 mice BT474 PTEN− LTT xenograft bearing mice before primary tumors reach 500 mm3. Data shown as average ± SD. *p ≤ 0.05, #p ≤ 0.01.

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