Fig. 5.

In vivo activation of PID by PDK1. PID was immunoprecipitated from cells cotransfected with HA-PID and Myc-PDK1 in the presence or absence of PDK1-RNAi. After PID autophosphorylation for the indicated times, MBP was added to the kinase reaction for an additional 30 min to asses PID transphosphorylation activity (Top). Equal amounts of proteins from the above transfected cells were used to confirm PID and PDK1 expression by immunoblotting by using HA (Middle) and Myc (Bottom) antibodies, respectively. Data are representative of two independent experiments.