Figure 4
TRAIL-R2-mediated apoptosis induction of scFv–scTRAIL. (a) 1 × 105 Jurkat cells/well were seeded in 96-well plates. The following day, cells were challenged with the indicated concentration of scFv–scTRAIL in the absence (open symbols) or presence (filled symbols) of 1 μg/ml of the crosslinking anti-FLAG mAb M2. After additional 16 h, cell viability was determined by MTT assay. (b) Cells (5 × 104 Colo205, 2 × 104 HT1080, 1 × 105 Jurkat cells/well) were seeded in 96-well plates. The next day, cells were treated with 2.5 μg/ml cycloheximide (HT1080, Colo205) and challenged with scFv–scTRAIL (1.35 nM) (HT1080, Colo205) or 2 nM TRAIL+1 μg/ml anti-FLAG mAb M2 (Jurkat) in the presence or absence of TRAIL-R2 Fab (50 μg/ml). After additional 16 h, cell viability was determined by MTT assay or KV staining. (c) 1 × 104 Colo205 and 1 × 104 L929 cells/well were seeded and co-cultured in 96-well plates. The next day, cells were treated with 2.5 μg/ml cycloheximide and challenged with 1 nM scTRAIL or scFv–scTRAIL. After additional 16 h, cell viability was determined by MTT assay. (a–c) Results from three independent experiments are shown (mean±S.D.)
