Figure 5.
PIP2 binding of IQGAP1 is important for cell morphology and migration. For both (A) and (B), Iqgap1 KO MEFs, reconstituted with the indicated proteins, were plated on 0.2% gelatin gel for 3 h. Fixed cells were stained for IQGAP1 and F-actin. Cells were photographed at × 600 magnification. (A) At least 300 cells were counted for each condition and categorized based on cell morphology (left). The graph is shown as mean of three independent experiments (right bottom). Expression levels of the proteins were analysed by immunoblotting with antibodies against the indicated molecules (right top). (B) IQGAP1 and F-actin staining. Arrowhead indicates the lamellipodium that is deficient of the ΔIQ mutant. (C) With the reconstituted MEFs, fibronectin-induced haptotaxis was measured as described in Figure 2E. (D) Reconstituted MEFs were plated on gelatin gel for 3 h before recording using time-lapse microscopy. Images were collected every min for 6 h at × 100 magnification and combined into a time-lapse movie. The migration path of six individual cells was then traced and plotted on a grid, with the origin of each cell placed in the centre of the grid. All the experiments described above were performed independently at least three times.
Source data for this figure is available on the online supplementary information page.
