Human Pak3-T421E cDNA with a flag tag was transfected into HEK293 cells using Lipofectomine 2000, After 72 hrs of incubation, PAK3 interactors were pulldwon with FLAG antibody. A sample without PAK3 overexpression was used as a negative control. Changes in PAK3 interactions between basal and starvation conditions were studied.