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. 2014 Sep 1;2(6):e00081. doi: 10.1002/prp2.81

Figure 5.

Figure 5

CD40 stimulation protects CLL cells from killing by TRAIL plus AZD5582. (A) CLL samples (n = 4) were cultured for 48 h on control or CD154-expressing fibroblasts in the presence or absence of AZD5582 (100 nmol/L) and then examined for expression of TRAIL receptor 1 (DR4) and 2 (DR5) by flow cytometry. Unstimulated CLL samples (n = 2) were also examined for basal levels of DR4 and DR5 (upper panel). Representative histograms are shown. (B) Quantification and statistical analysis of the data illustrated in A showing the effect of CD40 stimulation on TRAIL receptors 1 (DR4) and 2 (DR5). (C) CLL cells were cultured for 48 h on the respective fibroblasts monolayers in the presence or absence of AZD5582 (100 nmol/L). They were then removed from the cocultures, respectively, and incubated under standard conditions with or without TRAIL at the indicated concentrations for a further 24 h before being analysed for viability. (D) Unstimulated and cocultured CLL cells on control or CD154-expressing fibroblasts for 48 h as described in (C) were examined for level of cFLIP expression by Western blotting. β-actin was used as a loading control.