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Fig. 2 | Molecular Medicine

Fig. 2

From: Selective Inhibition of FLICE-like Inhibitory Protein (FLIP) Expression With Small Interfering RNA Oligonucleotides (siRNAs) Is Sufficient to Sensitize Tumor Cells for TRAIL-Induced Apoptosis

Fig. 2

FLIP-specific siRNAs sensitize tumor cells for TRAIL-induced apoptosis. SV80 and KB cells (107 cells/ml) were transiently transfected twice by electroporation on 2 consecutive days without (A) or with 150 nM of siRNA-neo (B), siRNA-F1 (C), siRNA-F2 (D), or a mixture of 75 nM each of siRNA-F1 and siRNA-F2 (E). In the first round of electroporation, a GFP expression plasmid was included in the electroporation mixture to verify transfection efficiencies. One day after the first electroporation, cells were harvested and analyzed for transfection efficiency by FACS analysis (solid line, first and third panel) or were electroporated a second time with the same siRNA as the first day. After the second electroporation cells were seeded in 100 µl medium in 96-well plates. The next day, cells were treated for 9 hr with Flag-tagged soluble TRAIL (50 ng/ml) cross-linked with anti-Flag mAb M2 (1 µg/ml) or agonistic TRAIL-R1- and TRAIL-R2-specific rabbit antisera (1:500), or remained untreated. Where indicated, cells were challenged with TRAIL in the presence of the caspase inhibitor z-VAD-fmk (20 µM). Finally, cell viability was determined using crystal violet staining (second and fourth panel).

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