Table 5. Lateral mobility of plasma membrane-bound GFP-tagged wildtype and mutant stomatin.
Effects of cholesterol and cytochalasin D.
| Stomatin mutant | Treatment | Mobile fraction (MF) | Recovery halftime (t1/2) | ||||||
|---|---|---|---|---|---|---|---|---|---|
| Mean (%) | SE | ΔMF (%) | p-value | Mean [s] | SE | Δt1/2 [s] | p-value | ||
| WT | none | 58.6 | 4.4 | ─ | ─ | 26.7 | 9.4 | ─ | ─ |
| WT | -chol | 53.9 | 7.7 | - 4.7 | 0.9217 | 41.4 | 16.7 | +14.7 | 0.1211 |
| WT | +chol | 57.9 | 2.7 | - 0.7 | 0.3673 | 29.4 | 4.2 | + 2.7 | 0.3573 |
| WT | +cytoD | 79.0 | 1.7 | +20.4 | 0.0022 | 18.8 | 2.5 | - 7.9 | 0.0330 |
| WT | -chol +cytoD | 61.3 | 3.8 | + 2.7 | 0.8306 | 33.8 | 11.9 | + 7.1 | 0.7506 |
| WT | +chol +cytoD | 69.1 | 2.1 | +10.5 | 0.1830 | 20.8 | 4.6 | - 5.9 | 0.1137 |
| ΔN | none | 60.3 | 2.4 | ─ | ─ | 44.5 | 8.3 | ─ | ─ |
| ΔN | -chol | 58.2 | 3.1 | - 2.1 | 0.8330 | 49.9 | 12.1 | + 5.4 | 0.3480 |
| ΔN | +chol | 59.3 | 3.1 | - 1.0 | 0.6186 | 28.9 | 13.5 | - 15.6 | 0.6259 |
| ΔN | +cytoD | 83.2 | 2.2 | +22.9 | <0.0001 | 25.6 | 1.4 | - 18.9 | 0.0007 |
| ΔN | -chol +cytoD | 70.7 | 5.4 | +10.4 | 0.0248 | 34.1 | 14.3 | - 10.4 | 0.7731 |
| ΔN | +chol +cytoD | 83.3 | 2.3 | +23.0 | <0.0001 | 17.7 | 3.0 | - 26.8 | 0.0003 |
| ΔC | none | 88.6 | 5.1 | ─ | ─ | 17.4 | 5.1 | ─ | ─ |
| ΔC | -chol | 85.7 | 2.8 | - 2.9 | 0.1971 | 22.2 | 2.1 | + 4.8 | 0.7227 |
| ΔC | +chol | 63.0 | 2.5 | - 25.6 | <0.0001 | 23.9 | 7.0 | + 6.5 | 0.2423 |
| ΔC | +cytoD | 88.6 | 3.0 | 0.0 | 0.5478 | 21.8 | 5.1 | + 4.4 | 0.9478 |
| ΔC | -chol +cytoD | 87.1 | 2.0 | - 1.5 | 0.2276 | 22.6 | 1.5 | + 5.2 | 0.7656 |
| ΔC | +chol +cytoD | 66.1 | 2.0 | - 22.5 | <0.0001 | 28.0 | 2.4 | +10.6 | 0.3684 |
| ΔCC | none | 69.6 | 1.5 | ─ | ─ | 29.1 | 3.5 | ─ | ─ |
| ΔCC | -chol | 75.4 | 2.4 | + 5.8 | 0.0407 | 26.5 | 3.3 | - 2.6 | 0.4965 |
| ΔCC | +chol | 64.9 | 2.6 | - 4.7 | 0.1844 | 25.9 | 5.9 | - 3.2 | 0.9956 |
| ΔCC | +cytoD | 83.5 | 3.4 | +13.9 | 0.0004 | 26.3 | 4.1 | - 2.8 | 0.5032 |
| ΔCC | -chol +cytoD | 77.7 | 2.3 | + 8.1 | 0.0120 | 19.9 | 2.6 | - 9.2 | 0.0175 |
| ΔCC | +chol +cytoD | 72.2 | 2.0 | + 2.6 | 0.4856 | 26.5 | 2.6 | - 2.6 | 0.2969 |
| Cys30Ser | none | 66.8 | 2.1 | ─ | ─ | 29.3 | 2.4 | ─ | ─ |
| Cys30Ser | -chol | 66.7 | 2.2 | - 0.1 | 0.9245 | 49.0 | 4.6 | +19.7 | 0.0003 |
| Cys30Ser | +chol | 55.8 | 3.7 | - 11.0 | 0.0897 | 26.0 | 5.4 | - 3.3 | 0.7465 |
| Cys30Ser | +cytoD | 81.1 | 1.9 | +14.3 | <0.0001 | 26.6 | 2.3 | - 2.7 | 0.5719 |
| Cys30Ser | -chol +cytoD | 73.0 | 1.5 | + 6.2 | 0.0392 | 37.3 | 1.8 | + 8.0 | 0.0137 |
| Cys30Ser | +chol +cytoD | 69.2 | 2.5 | + 2.4 | 0.4429 | 17.5 | 2.6 | - 11.8 | 0.0063 |
| Arg62Ala | none | 77.4 | 2.7 | ─ | ─ | 28.5 | 2.1 | ─ | ─ |
| Arg62Ala | -chol | 75.7 | 4.8 | - 1.7 | 0.5329 | 25.7 | 2.8 | - 2.8 | 0.3605 |
| Arg62Ala | +chol | 59.7 | 2.1 | - 17.7 | <0.0001 | 22.2 | 2.6 | - 6.3 | 0.0254 |
| Arg62Ala | +cytoD | 83.2 | 4.4 | + 5.8 | 0.1802 | 26.8 | 4.2 | - 1.7 | 0.5700 |
| Arg62Ala | -chol +cytoD | 85.4 | 2.7 | + 8.0 | 0.0289 | 26.1 | 3.3 | - 2.4 | 0.5237 |
| Arg62Ala | +chol +cytoD | 75.8 | 2.4 | - 1.6 | 0.7012 | 21.5 | 2.2 | - 7.0 | 0.0750 |
| Pro200Ala | none | 68.6 | 2.1 | ─ | ─ | 32.3 | 2.1 | ─ | ─ |
| Pro200Ala | -chol | 70.7 | 5.7 | + 2.1 | 0.1618 | 50.6 | 3.7 | +18.3 | 0.0002 |
| Pro200Ala | +chol | 62.8 | 3.0 | - 5.8 | 0.3068 | 24.5 | 7.7 | - 7.8 | 0.5346 |
| Pro200Ala | +cytoD | 76.3 | 1.2 | + 7.7 | 0.0089 | 19.3 | 1.3 | - 13.0 | 0.0009 |
| Pro200Ala | -chol +cytoD | 77.6 | 2.6 | + 9.0 | 0.0048 | 40.7 | 5.2 | + 8.4 | 0.0055 |
| Pro200Ala | +chol +cytoD | 72.5 | 1.9 | + 3.9 | 0.2533 | 28.6 | 2.2 | - 3.7 | 0.7598 |
| Pro245Ala | none | 78.0 | 2.7 | ─ | ─ | 34.5 | 2.4 | ─ | ─ |
| Pro245Ala | -chol | 61.8 | 2.1 | - 16.2 | 0.0005 | 36.0 | 6.1 | + 1.5 | 0.3403 |
| Pro245Ala | +chol | 66.7 | 5.2 | - 11.3 | 0.1293 | 30.9 | 3.5 | - 3.6 | 0.0279 |
| Pro245Ala | +cytoD | 78.6 | 2.3 | + 0.6 | 0.9900 | 21.2 | 1.3 | - 13.3 | <0.0001 |
| Pro245Ala | -chol +cytoD | 77.7 | 2.6 | - 0.3 | 0.9726 | 19.3 | 2.7 | - 15.2 | 0.0005 |
| Pro245Ala | +chol +cytoD | 74.0 | 1.9 | - 4.0 | 0.2923 | 24.8 | 3.4 | - 9.7 | 0.0695 |
MF, mobile fraction; t1/2, recovery half-time; ΔMF, MF-change of treated versus untreated sample; Δt1/2, t1/2-change of treated versus untreated sample; SE, standard error; WT, wildtype; ΔN, N-terminal deletion; ΔC, C-terminal deletion; ΔCC, coiled-coil deletion; chol, cholesterol; cytoD, cytochalasin D. Mean values, standard errors (SE), differences between mutants and WT values (ΔMF, Δt1/2), and p-values are given. Number of measurements for each value: n ≥ 20. The p-values indicate the significance of the differences between values of treated versus untreated samples.