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. 2016 Mar 7;7(16):21713–21727. doi: 10.18632/oncotarget.7964

Figure 3. Modulation of Notch expression and activation in AML and hBM-MSCs* upon co-culture.

Figure 3

A. Representative change in Notch1 level in AML cells: after 24 hours of co-culture with hBM-MSCs*, AML cells were collected and flow cytometry analysis for Notch1 expression was performed. The relative MFI was calculated and normalized as percentage with relative MFI of AML cells cultured alone. Data are expressed as mean ± SEM of 4 independent experiments. B. Representative western blot analysis of Hes1 in AML cells after 24 hours of co-culture with hBM-MSCs*. C. Representative RBP-jk activity in THP1 cells expressing RBP-jk-GFP reporter gene, THP1 cells were transfected with RBP-jk GFP reporter gene, after 4 hours cells were treated with Notch ligands or seeded on hBM-MSCs* monolayer. GFP levels were analyzed through flow cytometry after 72hours of co-culture. Data were normalized in percentage with THP1 transfected with CMV-GFP plasmid and are expressed as mean ± SEM of 4 independent experiments. D. Viability of primary leukemia cells cultured in presence of Notch ligands (Jagged1, Jagged2) or co-cultured with hBM-MSCs*. After 4 days, cells were collected and stained with Topro-3 to discriminate death cells through flow cytometry analysis. Data are representative of mean ± SEM of 3-7 independent experiments involving 3-10 patients.