Figure 5
Expression and function of Notch ligands in T-ALL cell lines.
(a) Expression of Notch ligands was analyzed with FACS in normal thymocytes and T-ALL cell lines. The ΔMFIs are shown. (b) Normal thymocytes (NT) (1 × 107) or T-ALL cells (1 × 106) were co-cultured with C2C12 cell monolayers in triplicate for 2 days and the transcripts of Notch1 and Jagged1 in C2C12 cells were determined with qRT-PCR. *; p < 0.05; **; p < 0.01. (c) FL0 cells were cultured in the absence or presence of DAPT and analyzed for the expression of Jagged1 with FACS. Shaded areas indicate the staining with control antibody. ΔMFIs at varying concentrations of DAPT are indicated. (d) T-ALL cell lines were cultured in the presence of control IgG or mixtures of monoclonal antibodies against Jagged1, Jagged2 and Dll1 (60 μg/mL each) for 1 day and immunoblotted with the indicated antibodies. Relative intensities of NICD to actin are indicated. Aliquots of the cells were cultured for 5 days and the viable cell numbers were determined in triplicate culture. (e) FL0 and T-lymphoma (EL4, RLm1) cell lines were cultured in methylcellulose medium (100 cells/dish) in triplicate for 7 days and the colony numbers were counted. Images of typical colonies of RLm1 and FL0 cells are shown. In (a), relevant parts of immunoblot images were cropped from full-length blots shown in Figure S10.
