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. 2015 Aug 4;290(39):23766–23780. doi: 10.1074/jbc.M115.673517

FIGURE 6.

FIGURE 6.

The IQGAP1-SKAP interaction links the MTs to cell cortex in cell migration. A, characterization of the efficiency and specificity of various siRNA oligonuclotides in MDA-MB-231 cells. B, in MDA-MB-231 cells, suppression of SKAP, IQGAP1, or SKAP/IQGAP1 exhibits the same defect in directional migration. Cells were treated with the indicated siRNAs for 72 h and scratched, followed by visualization with phase-contrast microscopy at the indicated time points. Scale bar, 100 μm. C, relative migration rates in B were calculated and graphed. Bars indicate S.D. from three independent experiments. Statistics significance was evaluated by an unpaired Student's t test. D, characterization of SKAP siRNA-resistant plasmids. Aliquots of MDA-MB-231 cells, transfected with three GFP-SKAP constructs, express at comparable levels judged by anti-GFP blotting analysis. E, real-time imaging of cell migration in MDA-MB-231 cells co-transfected with SKAP siRNA and the indicated SKAP siRNA-resistant plasmids. Migration tracks of transfected cells are shown as red lines. The migration paths of randomly picked transfected cells are presented for each group. Scale bars, 30 μm. F, relative migration speeds in E were calculated and graphed. Bars indicate S.D. from three independent experiments. Statistics significance was determined by an unpaired Student's t test.