Figure 8.

Rac1 RNAi differentially affect A-431 and A-431-C2β WT cell migration. (A and B) A-431 and A-431-C2β WT cells (Myc 6) cells were treated with or without EGF. (C) A-431 and A-431-C2β WT cells were transfected with Rac1 RNAi or scrambled control (−) and left for 48 h in culture to achieve target down-regulation. Cell migration tracks were monitored by live microscopy for 18 h, and migration speed (A and C) or horizon radius (B) was determined using Mathematica. These results are representative of a minimum of three independent experiments. Each condition was performed in quadruplicate using 15 cells per replicate. (D and E) Cell migration tracks of A-431 cells, or A-431 expressing kinase-dead PI3KC2β (C2β DN-17, C2β DN-32) were monitored by live microscopy for 18 h, and migration speed (D), or track paths (E) were determined using Mathematica. Results are representative of a minimum of three independent experiments. Each condition was performed in quadruplicate using 15 cells per replicate. (Significant difference as compared with control A-431 (A and D) or between Rac1 RNAi and scrambled control (C); Student’s t test: *p < 0.05; **p < 0.01; ***p < 0.001).