Fig. 3. TRAF6 induces the localization of PH-Akt-GFP to actin branch points. (A) HEK293 cells were co-transfected with a PH-Akt-GFP expression vector reporter and a TRAF6 expression vector for 24 hours, followed by fixation and phalloidin-TRITC staining. GFP (green), FITC (red) and merged images were used to determine signal co-localization (yellow). (B) HEK293 cells were co-transfected with PH-Akt-GFP and TRAF6 expression vectors for 24 hours, then treated for the indicated times with 5 µM cytochalasin D.