Fig. 3
GFP-tagged copines translocate to the plasma membrane in response to cAMP stimulation. Cells expressing GFP or a GFP-tagged copine were pulsed with 50 nM cAMP every 6 min for 5 h prior to plating cells in glass bottom dishes with 2 mM caffeine and 2 mM CaCl2. Cells were stimulated with cAMP and time-lapse images were obtained every 3 s after stimulation, time (seconds) of the image is noted in bottom right corner of each micrograph. Images were obtained with a Nikon Eclipse Ti inverted laser scanning confocal microscope using a 60× oil immersion objective. Representative images are shown. Scale bar = 5 μm. Time-lapse images were converted to movies at 2 frames per second using ImageJ (see Additional files 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12)