Fig. 2.

ACC coimmunoprecipitates with MIG12. (A) Five male mice were injected with an adenovirus expressing either β-gal or MIG12-FLAG. Two days after injection, mice were fasted for 12 h (F) or refed a high carbohydrate diet for 12 h following a 12 h fast (R). Livers were harvested and MIG12 was immunoprecipitated from liver lysates using an anti-FLAG monoclonal antibody. Cell lysates and proteins in immunoprecipitation pellets were separated by SDS/PAGE and immunoblot analysis was performed using polyclonal antibodies that recognize mouse ACC1, FAS, and ACL as described in Methods. (B) Five male mice were fed chow ad libitum (C), fasted for 12 h (F), or refed a high carbohydrate diet for 12 h after a 12 h fast (R). Livers were harvested and endogenous MIG12 was immunoprecipitated from cell lysates using an antimouse MIG12 polyclonal antisera or rabbit preimmune sera. Proteins in the supernatant and immunoprecipitation pellets were separated by SDS/PAGE and immunoblot analysis was performed using polyclonal antibodies that recognize mouse ACC1 and MIG12. * cross-reacting band.