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. 2012 May 25;2(5):e73. doi: 10.1038/bcj.2012.17

Figure 5.

Figure 5

Expression of IL-7R, VEGFR2, active caspase-3, Bcl-2, CD23, c-IAP protein, NF-κB and Hes-1 in CLL cells cocultured with hBM-MSCs. The expression of IL-7 R (a), VEGFR2 (b), active caspase-3 (c), CD23 (d), Bcl-2 (e), p53 (f) and NF-κB (g) by CD5+CD19+CD45+ CLL cells was analyzed by flow cytometry following coculture with hBM-MSCs (10/1 ratio), in absence or presence of anti-Notch-1,-2 or -4 antibodies or GSI XII for 3 days. Western blot analysis was performed to assess the expression of c-IAP2, Hes-1, PARP, NF-κB and Bcl-2 by CLL cells cocultured with hBM-MSCs in the same conditions (h). Hes-1 expression was used as functional read-out for Notch activation. NF-κB and Bcl-2 expression was evaluated to confirm CLL cell activation following coculture with hBM-MSCs in the same conditions. β-actin was included as loading control. Statistical analysis was carried out by using one-way analysis of variance, Holm–Sidak test. *P<0.05, **P<0.01, ***P<0.001 versus IgG-treated control were considered statistically significant. GSI XII was resuspended in dimethyl sulfoxide (DMSO) and DMSO was used as control for vehicle effects.