CIAR-PM-293 cells expressing Ras-LOCKR-S WT or NC localized to PM ( c ) or Golgi ( d ) were stimulated with either 100 ng ml −1 EGF or 250 nM A115. Left, representative epifluorescence images using ×60 objective (CFP channel from Key, FRET channel and pseudocolored raw FRET ratio) from three biologically independent experiments. Right, normalized FRET ratio changes ( n = 16 cells per condition for WT PM and NC Golgi experiments, n = 20 cells per condition for NC PM experiments and n = 15 cells per condition for WT Golgi experiments). Scale bars, 10 μm.
