Figure 8.

GFP-Ocrl1 lacking clathrin-/AP2-binding motifs or carrying non-functional phosphatase domain show decreased localization in membrane ruffles. (A andB) HT1080 cells expressing the indicated GFP-fusion proteins were starved for at least 8 h, stimulated for 15 min with 10% FBS, fixed and imaged. The number of cells displaying 0, 25, 50 or more than 75% of their perimeter decorated with ruffles positive for the corresponding GFP-fusion protein was determined. Values represent the mean ± SD of at least three independent determinations involving more than 100 cells randomly selected. Ruffling index (RI; see Materials and Methods) is also indicated. Representative images of each transfectant are included. Scale bars: 10 µm.