Table 1.
Values for podosome formation after various treatments
| Cells injected with | Cells with podosomes (%) | |
|---|---|---|
| 0 h | 1 h | |
| Mock | 8.9 ± 7.2 | 79.2 ± 11.0 |
| AMP-PNP (50 mM) | 36.7 ± 9.0 | |
| KIF5B antibody (4 μg/μl) | 72.2 ± 3.0 | |
| KIF3B antibody (4 μg/μl) | 78.9 ± 5.1 | |
| dynein antibody (4 μg/μl) | 80.0 ± 6.8 | |
| EHNA (1 mM) | 81.1 ± 3.9 | |
| Cells transformed with | 4 h | 6 h | 8 h |
|---|---|---|---|
| Kinesin construct (no.) | |||
| 1 (Control) | 93.3 ± 3.3 | 94.4 ± 6.9 | 87.8 ± 1.9 |
| 2 | 84.4 ± 5.1 | 83.3 ± 12.0 | 88.9 ± 1.9 |
| 3 | 55.6 ± 8.4 | 57.8 ± 11.7 | 72.2 ± 5.1 |
| 4 | 34.4 ± 6.9 | 50.0 ± 8.8 | 56.7 ± 15.3 |
| 5 | 71.1 ± 5.1 | 51.1 ± 7.7 | 47.8 ± 8.4 |
| 6 | 67.8 ± 10.2 | 54.4 ± 6.9 | 70.0 ± 6.7 |
| 7 | 63.3 ± 6.7 | 62.2 ± 6.9 | 76.7 ± 6.7 |
| 8 | 67.8 ± 10.2 | 56.7 ± 3.3 | 57.8 ± 10.7 |
| 9 | 63.3 ± 6.7 | 77.8 ± 5.1 | 65.6 ± 8.4 |
| 10 | 93.3 ± 5.8 | 93.3 ± 3.3 | 92.2 ± 5.1 |
| 11 | 97.8 ± 1.9 | 96.7 ± 3.3 | 96.7 ± 3.3 |
| 12 | 95.6 ± 1.9 | 95.6 ± 1.9 | 92.2 ± 1.9 |
| 13 | 92.2 ± 1.9 | 95.6 ± 1.9 | 91.1 ± 5.1 |
| 14 | 94.4 ± 1.9 | 91.1 ± 5.1 | 94.4 ± 1.9 |
| 15 | 95.6 ± 1.9 | 95.6 ± 1.9 | 94.4 ± 3.8 |
| 16 | 94.4 ± 3.8 | 96.7 ± 0.0 | 96.7 ± 0.0 |
| siRNA | 24 h | 48 h |
|---|---|---|
| Vector control (pEGFP-N1) | 78.0 ± 7.0 | 80.0 ± 7.0 |
| Luciferase | 74.0 ± 4.0 | 68.0 ± 17.0 |
| KIF1C | 40.0 ± 18.0 | 42.0 ± 2.0 |
| KIF3A | 63.3 ± 6.7 | 62.2 ± 6.9 |
| KAP3A | 67.8 ± 10.2 | 56.7 ± 3.3 |
| KIF5B | 54.4 ± 8.0 | 65.6 ± 11.0 |
| shRNA | |||
|---|---|---|---|
| Scrambled | 68.9 ± 5.9 | 65.6 ± 5.3 | |
| KIF1C | 32.2 ± 3.9 | 36.7 ± 5.1 |
| Dissolution (%) | Fission (%) | No change (%) | |
|---|---|---|---|
| Scrambled | 29.6 ± 9.0 | 30.6 ± 10.7 | 42.5 ± 3.9 |
| KIF1C | 23.1 ± 5.0 | 13.6 ± 7.4 | 63.0 ± 5.9 |
Statistical evaluation of podosome numbers or behavior in macrophages microinjected with inhibitors/inhibitory antibodies, transformed with various kinesin constructs, or transformed with siRNA or shRNA. For each value, each time 30 randomly chosen cells from three independent experiments were evaluated. For evaluation of podosome behavior, a total of 473 podosomes for scrambled shRNA and a total of 501 podosomes for KIF1C shRNA from each time three independent experiments were evaluated. Values are given as mean percentage ± SD of total counts (see Figure 2).