Research Paper Volume 12, Issue 14 pp 15104—15120

Figure 6. STAT5a overexpression in human DCIS cells drives invasion, a phenomenon enhanced by 17-β-Estradiol treatment. (A) Qualitatively, transwell invasion assay results depict DAPI-stained invaded cells after 18- hour treatment of MCF10DCIS.com empty vector or STAT5a overexpressor cells with vehicle (DMSO) or 1nM estradiol. Images of the invaded cells were captured using the DAPI channel on the EVOS FL microscope at 10x objective. (B) Quantitatively, the number of invaded cells in five representative fields of view were averaged for each membrane. Data were normalized to empty vector. Treatment with vehicle or 1nM estradiol led to a significant increase in invasion of STAT5a overexpressor cells compared to empty vector cells (vehicle-treated empty vector vs. STAT5a overexpressor cells: 1.9-fold, p<0.001, n=3; estrogen-treated empty vector vs. STAT5a overexpressor cells: 2.4-fold, p<0.001, n=3). Moreover, the invasion of STAT5a overexpressor cells relative to empty vector cells was calculated independently for each treatment group and reported as a median change in invasion with an associated interquartile range (IQR) (vehicle [STAT5a overexpressor/empty vector]: 1.95 median, 0.49 IQR, p<0.001, n=3; estrogen [STAT5a overexpressor/empty vector]: 2.54 median, 0.39 IQR, p<0.001, n=3). The median change in invasion of estrogen-treated STAT5a overexpressor cells relative to empty vector cells was significantly higher compared to the median change in invasion of vehicle-treated STAT5a overexpressor cells relative to empty vector cells (p<0.001, n=3).