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Figure 2 | BMC Cancer

Figure 2

From: Tissue factor/FVIIa activates Bcl-2 and prevents doxorubicin-induced apoptosis in neuroblastoma cells

Figure 2

The effects of TF silencing by siRNA on the expression of endogenous TF, Bcl-2 and Bcl-XL as well as cell growth and apoptosis induced by doxorubicin in SH-EP1. A, cells cultured with 10 nM FVIIa were treated with different concentrations of either TF siRNA or control siRNA for 24 h. Expression of endogenous TF, Bcl-2 and Bcl-XL was detected by western blot assay. B, clonogenic assay of SH-EP1 in culture with TF siRNA. Cells (1 × 104) were cultured in soft agarose in the presence of FVIIa for 2 weeks with different doses of TF siRNA and control siRNA, respectively, and resulting colonies counted. Data for the colony counts is shown (mean ± S.D. for triplicate cultures). C, western blot assay for activation of caspase-3 and cleavage of the death substrate PARP in SH-EP1 cells that were treated with 2 μM doxorubicin (Dox) and 200 nM of either TF siRNA or control siRNA for the indicated time points. D. quantitative detection of apoptotic cells induced by Dox when given in combination with either TF siRNA or control siRNA. SH-EP1 cells were treated with Dox (2 μM) plus siRNA (200 nM) for the indicated times, and apoptotic cells were detected by flow cytometry. Data represents the percentage of annexin-V positive cells.

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