FIG. 13.

PRMPs spatially correlate with Myo1c-induced membrane ruffles. Ultrafast-acquisition deconvolution microscopy was carried out every 30 s for 10 min for a living adipocyte expressing both ECFP/PLCδ1-PH and EYFP/Myo1c fusions. The CFP fluorescence was excited with the 458-nm laser line of an Ar laser. Therefore, there is significant fluorescence bleedthrough from the CFP channel to the YFP channel, while reverse bleedthrough is negligible. Therefore, only the CFP fluorescence images depicting PtdIns(4,5)P₂ distribution and dynamics are shown, with the understanding that major membrane ruffles (labeled 1 to 3 in panel A) are caused by Myo1c overexpression (see Results). Deconvolved images were projected within 3D grids measuring 18 by 42 by 10 μm and were visualized with 8-bit false-color intensity scales (rescaled at between 10 and 100). Representative images at 0, 2, 4, 6, 8, and 10 min are shown. O, origin. (See movie 8 at http://invitro.umassmed.edu/∼sh/supplements/supplements.html for the complete image sequence.)