Figure 3
From: CD70-restricted specific activation of TRAILR1 or TRAILR2 using scFv-targeted TRAIL mutants
Apoptosis induction by TRAILR1 and TRAILR2 discriminating TRAIL mutants. (a) OVCAR3, Mino and Jurkat cells were analyzed with respect to cell surface expression of TRAILR1 and TRAILR2 by FACS. (b) OVCAR3, Mino and Jurkat cells were challenged in triplicates with the indicated concentrations of Flag-TNC-TRAIL, Flag-TNC-TRAILmutR1 and Flag-TNC-TRAILmutR2 in the presence and absence of 1 μg/ml of the anti-Flag mAb M2. Next day, cellular viability was determined using the MTT assay or crystal violet staining. OVCAR3 cells were challenged in the presence of 2.5 μg/ml CHX, which sensitizes this cell line for apoptosis induction. (c) The indicated cell lines were treated for 4–6 h with 100 ng/ml of the various Flag-TNC-TRAIL variants oligomerized with 1 μg/ml of the anti-Flag mAb M2, and total cell lysates were assayed by western blotting for the presence of the indicated proteins. OVCAR3 cells were again sensitized for apoptosis induction by CHX treatment
