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2: Alix co-immunoprecipitates with DN-pro-caspase-8 ,
ALG-2 is detected in pro-caspase-8/ Alix co-immunoprecipitates. BHK cells co-expressing Flag-Alix together with HA-DN-pro-caspase-8 were lysed and immunoprecipitated as in B; immunoprecipitates were analysed by Western Blot (WB) with polyclonal anti HA , anti-ALG-2 and anti-Flag antibodies ,
Alix with DN-pro-caspase-8 requires its ALG-2 binding domain. BHK cells were co-transfected with either Flag-Alix WT or Flag-AlixALG-2 and HA-DN- pro-caspase-8. IP with anti-HA antibody were performed in CaRIPA, Immunoprecipitates were analysed by WB using polyclonal antibodies against HA (upper) and Alix ,
RIPA lysates of HEK 293 cells co-expressing Flag-Alix and TNF-R1 were immunoprecipitated with anti-Flag antibodies and blots probed with anti-Alix (upper) ,