English: Representative FRET movie of a control siRNA-treated cell. HeLa cells treated with control (CTRL) siRNA were transfected with YFP-AktPH and CFP-AktPH. Transfected siRNA-treated cells were placed on the microscope stage at 37°C and image series were collected every 15 seconds for 15 minutes. Purified InlB was added to a final concentration of 5 nM after frame 15 (i.e. time point just prior to 4 minutes) after the start of imaging. A movie for the entire timecourse of InlB stimulation for the control cell depicted in Figure 5A is here presented (i.e. frame 0 to 60), showing the collected images (phase-contrast, Akt recruitment, and FRET efficiency). Pseudocolor scale represents the range of FRET efficiency values from original signal ranging from blue (low) to red (high).