Impact of SKAP55 mutants on SLP-76 microcluster dynamics
| SKAP55 chimera | Persistence | P | Inward movement | P | Max speed | P | % SLP-76 in clusters | P | Clusters tracked per diameter | P | Totalcells | n (experiments) |
| s | µm | nm/s | ||||||||||
| mRFP1 | 210.5 ± 7.20 | 3.0 ± 0.2 | 147.7 ± 6.1 | 62.0 ± 2.8 | 15.1 ± 0.48 | 46 | 19 | |||||
| KD | 115.8 ± 38.2 | ** | 0.6 ± 0.1 | ** | 57.8 ± 8.6 | ** | 27.3 ± 6.9 | ** | 12.5 ± 1 | *, †† | 10 | 3 |
| WT | 215.7 ± 7.52 | 3.4 ± 0.3 | 150 ± 8.6 | 72.1 ± 2.3 | 17.2 ± 0.87 | * | 29 | 15 | ||||
| WR | 129.3 ± 18.9 | **, †† | 1.3 ± 0.3 | **, †† | 127.8 ± 31.9 | 37.6 ± 6.6 | **, †† | 13.6 ± 1.8 | 8 | 3 | ||
| SH3 | 150.5 ± 16.4 | **, †† | 1.4 ± 0.5 | **, †† | 104.6 ± 23.4 | *, † | 41.9 ± 15.4 | *, †† | 9.6 ± 1.5 | **, †† | 10 | 5 |
| ΔPH | 222.9 ± 2.1 | 2.8 ± 0.2 | 154.2 ± 9.6 | 77.3 ± 2.9 | 15.3 ± 0.33 | 10 | 3 | |||||
| 3YF | 169 ± 16.4 | *, †† | 2.3 ± 0.3 | 153.5 ± 17.4 | 66.5 ± 5.1 | 16.4 ± 0.98 | 10 | 4 | ||||
| ΔDM | 175.9 ± 23.3 | 1.5 ± 0.2 | **, †† | 137.7 ± 15.1 | 44.9 ± 5.2 | **, †† | 13.3 ± 1.5 | † | 10 | 3 | ||
| DM | 133.1 ± 5.9 | **, †† | 1.1 ± 0.1 | **, †† | 101.6 ± 6.2 | **, † | 32 ± 4.0 | **, †† | 14 ± 2 | 10 | 3 | |
| TD | 227.2 ± 11.6 | 2.5 ± 0.2 | 129.1 ± 23.8 | 73.1 ± 1.1 | 14 ± 0.95 | † | 10 | 3 |
| SKAP55 chimera | Persistence | P | Inward movement | P | Max speed | P | % SLP-76 in clusters | P | Clusters tracked per diameter | P | Totalcells | n (experiments) |
| s | µm | nm/s | ||||||||||
| mRFP1 | 210.5 ± 7.20 | 3.0 ± 0.2 | 147.7 ± 6.1 | 62.0 ± 2.8 | 15.1 ± 0.48 | 46 | 19 | |||||
| KD | 115.8 ± 38.2 | ** | 0.6 ± 0.1 | ** | 57.8 ± 8.6 | ** | 27.3 ± 6.9 | ** | 12.5 ± 1 | *, †† | 10 | 3 |
| WT | 215.7 ± 7.52 | 3.4 ± 0.3 | 150 ± 8.6 | 72.1 ± 2.3 | 17.2 ± 0.87 | * | 29 | 15 | ||||
| WR | 129.3 ± 18.9 | **, †† | 1.3 ± 0.3 | **, †† | 127.8 ± 31.9 | 37.6 ± 6.6 | **, †† | 13.6 ± 1.8 | 8 | 3 | ||
| SH3 | 150.5 ± 16.4 | **, †† | 1.4 ± 0.5 | **, †† | 104.6 ± 23.4 | *, † | 41.9 ± 15.4 | *, †† | 9.6 ± 1.5 | **, †† | 10 | 5 |
| ΔPH | 222.9 ± 2.1 | 2.8 ± 0.2 | 154.2 ± 9.6 | 77.3 ± 2.9 | 15.3 ± 0.33 | 10 | 3 | |||||
| 3YF | 169 ± 16.4 | *, †† | 2.3 ± 0.3 | 153.5 ± 17.4 | 66.5 ± 5.1 | 16.4 ± 0.98 | 10 | 4 | ||||
| ΔDM | 175.9 ± 23.3 | 1.5 ± 0.2 | **, †† | 137.7 ± 15.1 | 44.9 ± 5.2 | **, †† | 13.3 ± 1.5 | † | 10 | 3 | ||
| DM | 133.1 ± 5.9 | **, †† | 1.1 ± 0.1 | **, †† | 101.6 ± 6.2 | **, † | 32 ± 4.0 | **, †† | 14 ± 2 | 10 | 3 | |
| TD | 227.2 ± 11.6 | 2.5 ± 0.2 | 129.1 ± 23.8 | 73.1 ± 1.1 | 14 ± 0.95 | † | 10 | 3 |
SLP-76 microcluster dynamics in J14.SY cells expressing each of the indicated constructs were measured on a per cell basis by manual tracing of microcluster trajectories from SLP-76 kymographs. Values for each experiment were obtained by averaging these per cell values, weighing each cell equally. KD, knockdown. Values displayed in the table are mean ± SEM for the indicated number of experiments. From parental J14.SY cells expressing mRFP1: *, P < 0.05; **, P < 0.01. From J14.SY cells expressing SKAP55.WT.mRFP1: †, P < 0.05; ††, P < 0.01.