Figure 3.

TNF-α-induced cell death in EVER2-expressing cells is mediated by interaction of EVER2 TMC domain with TRADD. (a) Schematic representation of the putative EVER2 domains and fragments used. TM: transmembrane domain; TMC: transmembrane conserved domain; FL: full-length; d1–d7: EVER2 fragments. (b) Schematic representation of the TRADD fragments used. FL: full-length; d1–d2: TRADD fragments. (c) Mapping of the interaction of EVER2 with TRADD fragments by HT–GPCA. (d) Mapping of the interaction of TRADD with EVER2 fragments by HT–GPCA. (e) Impact of EVER2 fragments on TNF-α-induced apoptosis. Cells were transfected with constructs encoding the various EVER2 fragments and TNF-α-induced apoptosis was assayed as described above. (f) Detection of EVER2 fragments expression in transfected HEK-293T using western blot analysis. Asterisks indicate the band corresponding to the expected molecular weight of each EVER2 fragment. Means±S.E.'s (n=3 separate experiments) are shown. The effect of the full-length protein was taken as the reference in panels D and E. Student's t-test comparing effect of EVER2 FL with that of other EVER2 fragments: ***P<0.001; **P<0.005; *P<0.01