Fig. 4. In contrast to its activity in the Wnt pathway, XIAPT180A has a similar activity to that of wild-type XIAP in the apoptotic pathway. (A) Smac co-immunoprecipitates (IP) with XIAP and XIAPT180A to a similar extent. HEK293STF cells were transfected as indicated with vector or HA–Smac plus MYC–XIAP or MYC–XIAPT180A. Lysates were collected and immunoprecipitation performed with anti-MYC antibody followed by immunoblotting with anti-HA antibody. WCL, whole cell lysates. Results were replicated at least three times. (B) Overexpression of XIAP and XIAPT180A decreases the percentage of Fas ligand (FasL)-induced apoptosis to a similar extent. HeLa cells were transfected for 24 h as indicated and incubated in the absence or presence of recombinant Fas ligand (100 ng/ml) for 24 h. Cells were then stained with Annexin V and propidium iodide. Graph shows the mean±s.d. of percentage of apoptotic cells. Data was analyzed and processed using FlowJo. **P<0.001, *P<0.01; n.s., not significant (one-tailed Student's t-test). All experiments were repeated at least three times. (C) A model for the regulation of XIAP activity in the Wnt pathway by phosphorylation at position T180. See text for more information.
Image published in: Ng VH et al. (2018)
Copyright © 2018. Image reproduced with permission of the Publisher, The Company of Biologists Ltd.
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