Fig. 3
FAF1 interacts with JNK1 upon ischemic stress. a HEK 293 T cells were transfected with the indicated combinations of HA-FAF1 (1 μg) and Flag-JNK1 plasmids (1 μg). At 48 h after transfection, the cells were untreated or treated with oxygen glucose deprivation for 30 min. Whole cell lysates were immunoprecipitated with anti-Flag antibody, followed by immunoblotting with the indicated antibodies. b Faf1+/+ MEFs were treated with oxygen glucose deprivation for the indicated times. The cell lysates were then immunoprecipitated with anti-JNK1 antibody. The interaction was determined by immunoblotting with the indicated antibody. Filled triangle denotes a non-specific band (c) Upper panel: GST pull-down assay in which in vitro-translated 35S-labeled JNK1 was incubated with GST and GST-FAF1 fragments, and the in vitro-translated proteins present in the input (5%) or those associated with FAF1 were detected by autoradiography. Lower panel: Schematic diagram of full length and truncated FAF1 constructs. FID, Fas-interacting domain; DEDID, death effecter domain-interacting domain; C-term, c-terminal domain