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. 2013 Feb 11;210(2):321–337. doi: 10.1084/jem.20121527

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© 2013 Kannan et al.

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Figure 7. — Notch ligand DLL1 co-culture inhibits AML growth, induces apoptosis, down-regulates BCL2, and reveals the critical roles of HES1, BCL2, and p53 in Notch-mediated apoptosis in AML. (A) Cell counts after 72 h of co-culture on human stromal line HS5 transduced with GFP-only or Notch ligand DLL1 GFP. 100,000 AML or T-ALL cells were placed over confluent HS5-GFP or HS5-DLL1 cells. (**, P < 0.001). Triplicate samples analyzed. (B) Flow cytometry–based apoptosis assay in AML and T-ALL cells stained with Annexin V after co-culture with HS5-GFP or HS5-DLL1 cells for 24 h. **, P < 0.001. Triplicate samples were analyzed. (C) Immunoblot of AML and T-ALL cells after co-culture with HS5-GFP or HS5-DLL1 cells for 48 h. Representative of two blots. (D) mRNA expression of BCL2 in AML and T-ALL co-cultured with HS5-GFP or HS5-DLL1 cells for 24 h. **, P < 0.001. Triplicate samples analyzed. (E) Flow cytometry–based apoptosis assay of AML cells transfected with siRNA to HES1 or control, and then co-cultured on HS5-GFP or HS5-DLL1 for 48 h. Representative of two experiments. (F) Immunoblot showing effect of transfection with siRNA to HES1 on level of HES1 protein induced by transduction with ICN1. Representative of two blots. (G) Flow cytometry–based apoptosis assay of AML cells transfected with BCL2, siP53, or BCL2 + siP53 was then co-cultured on HS5-GFP or HS5-DLL1 for 48 h. Representative of two experiments.