Figure 4
From: EVER2 protein binds TRADD to promote TNF-α-induced apoptosis
TNF-α induces caspase-8-dependent apoptosis in EVER2-expressing cells, by facilitating FADD/RIPK1/caspase-8 complex II formation. (a) TNF-α-induced apoptosis in EVER2-expressing cells is impaired by the inhibition of the activity of caspases . HEK-293T cells were transfected with EVER2–Flag or empty vector. Cells were stimulated with TNF-α, together with z-VAD or z-IETD. Values are means±S.E.’s (n=3). Student’s t-test: ***P<0.001. (b) The cleavage of caspase-8, caspase-3 and PARP was observed after 3 h of TNF-α stimulation in EVER2-expressing cells. Cells were transfected with the EVER2–Flag construct or empty vector. Cells were stimulated with TNF-α for various time periods. Immunoblotting was performed on whole-cell lysates with the indicated antibodies. (c) Profile of apoptosis-regulator protein levels during TNF-α stimulation in EVER2-expressing cells. Cells were treated as previously described and immunoblotting was performed on whole-cell lysates with the indicated antibodies. (d) FADD/RIPK1/caspase-8 complex II formation in EVER2-expressing cells. Cells were transfected with the EVER2–Flag construct or empty vector. Cells were stimulated with TNF-α (10 ng/ml) for various time periods, lysed and subjected to IP with a specific antibody against caspase-8. IB was performed with the indicated antibodies
