Minimal Cross-inhibition by Dominant Negative Inhibitors
| Transfectant . | Additional plasmid . | Migration rate ± SD . | Directionality . |
|---|---|---|---|
| μm/3 h | D/T ratio | ||
| PTEN + Shc | None | 110 ± 24 | 0.57 ± 0.20 |
| PTEN + Shc | FRNK | 101 ± 20 | 0.53 ± 0.25 |
| PTEN + Shc | Dn-Cas | 105 ± 18 | 0.50 ± 0.20 |
| PTEN + FAK | None | 117 ± 24 | 1.72 ± 0.14 |
| PTEN + FAK | Dn-Shc | 104 ± 21 | 1.81 ± 0.16 |
| PTEN + Cas | None | 118 ± 23 | 1.65 ± 0.17 |
| PTEN + Cas | Dn-Shc | 108 ± 18 | 1.82 ± 0.18 |
| Transfectant . | Additional plasmid . | Migration rate ± SD . | Directionality . |
|---|---|---|---|
| μm/3 h | D/T ratio | ||
| PTEN + Shc | None | 110 ± 24 | 0.57 ± 0.20 |
| PTEN + Shc | FRNK | 101 ± 20 | 0.53 ± 0.25 |
| PTEN + Shc | Dn-Cas | 105 ± 18 | 0.50 ± 0.20 |
| PTEN + FAK | None | 117 ± 24 | 1.72 ± 0.14 |
| PTEN + FAK | Dn-Shc | 104 ± 21 | 1.81 ± 0.16 |
| PTEN + Cas | None | 118 ± 23 | 1.65 ± 0.17 |
| PTEN + Cas | Dn-Shc | 108 ± 18 | 1.82 ± 0.18 |
U-87MG cells were cotransfected with PTEN plus the indicated plasmids, and then analyzed by video time-lapse microscopy and image analysis for rates of cell migration and directionality as described in Materials and Methods. Dn-Cas, dominant negative Cas plasmid ΔSD-p130Cas lacking the substrate domain. Dn-Shc, dominant negative Shc plasmid Y239/317F with a double mutation in tyrosine phosphorylation sites. These dominant interfering plasmids substantially inhibited migration in cells in which the FAK and Shc pathways were not inhibited by PTEN (see Fig. 2).