Figure 2.

Immunoblotting and Luciferase Reporter Assay of Notch1^high (N1ICD), Notch1^low (dnCSL), or native MDA-MB-231 breast cancer cells. (A) MDA-MB-231 breast cancer cells transfected with ΔEN1ICD or dnCSL show that ΔEN1ICD increases the levels of N1ICD protein, while no changes were detected in dnCSL as compared to native MDA-MB-231 cells. Anti-β-actin antibody was used as a control for equal loading. Representative blots from tree-independent experiments are shown; (B) Notch signaling activity from the 12xCSL-luc reporter in MDA-MB-231, Notch1^lowMDA-MB-231, and Notch1^highMDA-MB-231 breast cancer cells. The relative activity of Notch1 was normalized to the activity of native MDA-MB-231 cells. The results are presented as mean ± standard error of mean (±S.E.M). Statistical analysis was performed using a one-way ANOVA test, Tukey post-hoc testing (*** p < 0.001).