Fig. 8. AS160 becomes phosphorylated in the Rip11 complex in response to insulin. CHO.T cells were cotransfected with plasmids encoding FLAG-tagged wild-type AS160 (AS160-WT) together with either Xpress-tagged wild-type Rip11 (Rip11-WT) or Rip11-I630E, as shown. The cells were incubated in the absence or presence of 100 nM insulin for 10 minutes, as indicated, before lysis and immunoprecipitation of Rip11 complexes with antibodies against Xpress. These complexes were blotted with the antibody against FLAG (top panel) or the antibody against PAS, which recognises the PKB phosphorylation consensus sequence(s) present on AS160 (lower panel).