S1P release from apoptotic cells
| Sample . | S1P, mean ng/mL (n ≥ 4) . | SEM . |
|---|---|---|
| Jurkat cells, ×106 cells/mL | ||
| Control | 0.23 | ± 0.02 |
| 2 h staurosporine (0.5 ng/mL) | 0.29 | ± 0.02 |
| 4 h staurosporine (0.5 ng/mL) | 0.41 | ± 0.02 |
| 6 h staurosporine (0.5 ng/mL) | 1.11 | ± 0.09 |
| EGTA + 6 h staurosporine (0.5 ng/mL) | 0.33 | ± 0.04 |
| Ac-YVAD-CMK (1μM) + 6 h staurosporine (0.5 ng/mL) | 0.36 | ± 0.03 |
| HEK293 cells, 2 × 105 cells/mL | ||
| Control | ND | ND |
| 12 h staurosporine (0.5 ng/mL) | 0.2 | ± 0.02 |
| HA-SphK2 + 12 h staurosporine (0.5 ng/mL) | 5.37 | ± 0.51 |
| HA-SphK2-D138A + 12 h staurosporine (0.5 ng/mL) | 0.49 | ± 0.05 |
| HA-SphK2-T219A + 12 h staurosporine (0.5 ng/mL) | 0.22 | ± 0.03 |
| Sample . | S1P, mean ng/mL (n ≥ 4) . | SEM . |
|---|---|---|
| Jurkat cells, ×106 cells/mL | ||
| Control | 0.23 | ± 0.02 |
| 2 h staurosporine (0.5 ng/mL) | 0.29 | ± 0.02 |
| 4 h staurosporine (0.5 ng/mL) | 0.41 | ± 0.02 |
| 6 h staurosporine (0.5 ng/mL) | 1.11 | ± 0.09 |
| EGTA + 6 h staurosporine (0.5 ng/mL) | 0.33 | ± 0.04 |
| Ac-YVAD-CMK (1μM) + 6 h staurosporine (0.5 ng/mL) | 0.36 | ± 0.03 |
| HEK293 cells, 2 × 105 cells/mL | ||
| Control | ND | ND |
| 12 h staurosporine (0.5 ng/mL) | 0.2 | ± 0.02 |
| HA-SphK2 + 12 h staurosporine (0.5 ng/mL) | 5.37 | ± 0.51 |
| HA-SphK2-D138A + 12 h staurosporine (0.5 ng/mL) | 0.49 | ± 0.05 |
| HA-SphK2-T219A + 12 h staurosporine (0.5 ng/mL) | 0.22 | ± 0.03 |
S1P contents in supernatants were measured by liquid chromatography tandem mass spectrometry as described in “S1P quantification in cell-culture supernatants using LC-MS/MS.” The reliable lower limit of quantification was 0.2 ng/mL. ND indicates S1P levels below this limit.