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Figure 6 | Breast Cancer Research

Figure 6

From: Nicastrin and Notch4 drive endocrine therapy resistance and epithelial to mesenchymal transition in MCF7 breast cancer cells

Figure 6

Stable Nicastrin (NCST) overexpression confers 4-OH-TAM resistance in MCF7 cells through Notch4 activation. (A) MCF7 CT and MCF7 NCST cells treated with vehicle (EtOH) or 10-7 M 4-OH-TAM were plated (3 × 103/well) in 96-well plates and allowed to adhere. One plate was fixed and annotated as Day 0. A sulforhodamine B (SRB) assay was performed every two days until Day 6. The experiment was repeated three times and each time six technical replicates were used. (B) Representative western blot of estrogen receptor alpha (ERα) protein levels in MCF7 CT and MCF7 NCST cells. ActinB was used as loading control. (C) Quantification of microRNA (mRNA) levels of ERα target genes analysed by qRT-PCR. Fold change is shown in MCF7 NCST compared to MCF7 CT cells, normalised to GAPDH. Results represent three biological as well as three technical replicates of each. (D) Representative western blot confirming Notch4 small interfering RNA (siRNA) in MCF7 NCST cells and the effect on NCST protein. (E) An SRB assay performed as in A following siRNA transfection in MCF7 NCST. The experiment was repeated three times and each time six technical replicates were used.

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