FIG. 1.

Interaction of V and Akt1. (a) Coimmunoprecipitation of V and Akt1. HeLa cells were transfected with vector control (empty vector) or plasmids encoding V and/or Akt1. After 24 h, transfected cells were metabolically labeled with [³⁵S]Cys and [³⁵S]Met, lysed, and then subjected to immunoprecipitation with anti-V or anti-Akt1 antibody. The precipitates were resolved by 15% SDS-PAGE, and proteins were visualized by phosphorimagery. Migration of V and Akt1 are indicated on the right, and coprecipitated V or Akt1 are marked with asterisks. (b) BiF analysis of V and Akt1 interaction. HeLa cells were transfected with the following combinations of plasmids: YFP1-Zip and YFP2-V (negative control), YFP1-Zip and YFP2-Zip (positive control), or YFP2-V and YFP1-Akt1. Cells were collected, and fluorescence was measured by flow cytometry. The region considered YFP positive is indicated.