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. 2003 Feb;23(4):1428–1440. doi: 10.1128/MCB.23.4.1428-1440.2003

FIG. 5.

FIG. 5.

FasL and ACRP:FasL bind Fas equally well. (A) The interaction between Flag-tagged FasL, ACRP:FasL, and ACRPΔ:FasL on the one hand and coated Fas:Fc (filled symbols) or TRAILR2:Fc (open symbols, negative control) on the other hand was monitored by ELISA. (B) Burkitt lymphoma Raji cells were incubated for 15 min with FasL, ACRP:FasL, or PBS alone. Washed cells were lysed and FasL bound to the cells was detected by anti-Flag Western blotting. Cross-linking antibody was added during the incubation (+) or after cell lysis (−). (C) The binding of [125I]-labeled ZB4 anti-Fas antibody to BJAB cells was competed with the indicated concentrations of recombinant FasL (squares) or ACRP:FasL (circles). (D) The binding of Flag-tagged FasL or ACRP:FasL (at 200 ng/ml) to recombinant Fas:Fc was competed with the indicated amounts of untagged Fc:FasL. Bound ligands were detected via the Flag tag. (E) HeLa cells were incubated for 16 h with a constant, lethal concentration of ACRP:FasL in the presence of various amounts of FasL. Cell viability was monitored with the PMS/MTS reagent.