FIG. 5.
PI-3,4,5-P3 is generated at the plasma membrane as well as within cellular structures formed by actin remodeling. L6 myoblasts were transiently transfected with 0.5 μg of plasmid encoding GFP-PH-GRP1 and stimulated with insulin following 4 h of serum starvation. The cells were fixed and stained for (A) F-actin using rhodamine-phalloidin or (B) the plasma membrane using tetramethylrhodamine-conjugated ConA (50 μg/ml). Optical sections of 0.2 μm from the top of the cell to the bottom were acquired and deconvolved. A cross-section of the structure was obtained (a) and analyzed for colocalization with F-actin (Ab) or the plasma membrane (Bb) by selecting for yellow regions.