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. 2020 Oct 12;11:5129. doi: 10.1038/s41467-020-18860-4

Fig. 4. Endothelial Jag1-induced activation of Notch1 contributes to CSC phenotypes through a Zeb1-dependent mechanism.

Fig. 4

a, b Relative mRNA (a) and protein (b) levels of Zeb1 in MDA-MB-231 cells that were cocultured with shJag1- or shDll4-transfected HUVEC-mCherry. c, d Relative mRNA (c) and protein (d) levels of Zeb1 in shNotch1-, shNotch2- or shNotch3-transfected MDA-MB-231 cells that were cocultured with HUVEC-mCherry. e–g Flow cytometry analysis of e the CD44+CD24− population, f the ALDH activity, and g the side-population assay for shCtrl- or shZeb1-transfected MDA-MB-231 cells that were cocultured with HUVEC-mCherry. h Protein levels for Zeb1, Sox2, Oct4, and Nanog in shCtrl- or shZeb1-transfected MDA-MB-231 cells that were cocultured with HUVEC-mCherry. i In vivo tumorigenicity assay with limited dilution using shCtrl- or shZeb1-transfected MDA-MB-231 cells that were cocultured with HUVECs. The estimated CSC frequency was analyzed using ELDA software. Indicated P-values were calculated using two-tailed unpaired Student’s t-test in a, c, e–g, or chi-square test with multiple comparisons in i. Data are presented as mean ± SEM in a, c, e–g. Data are representative of three (a–h) independent experiments. Source data are provided as a Source Data file.