Table 3. Effect of TKTL1 knock down on sphingolipid synthesis in THP-1 cells.
| Species | Chain length (R) | THP-1WT | THP-1KD |
|---|---|---|---|
| Ceramides (Cer) | 14:1 | 2.43 ± 0.3 | 0.89 ± 0.2 |
| 14:0 | 8.40 ± 1.2 | 3.28 ± 1.1 | |
| 16:1 | 8.40 ± 1.5 | 2.91 ± 0.8 | |
| 16:0 | 158 ± 23 | 107 ± 40 | |
| 18:1 | 3.99 ± 0.3 | 1.47 ± 0.3 | |
| 18:0 | 20.8 ± 3.0 | 11.8 ± 5.2 | |
| 20:1 | 0.82 ± 0.12 | 0.30 ± 0.12 | |
| 20:0 | 4.67 ±0.4 | 2.59 ± 1.3 | |
| 22:1 | 4.52 ± 0.2 | 2.01 ±0.5 | |
| 22:1● | 7.30 ± 1.0 | 2.71 ± 0.5 | |
| 22:0 | 40.9 ± 5.7 | 25.9 ± 6.3 | |
| 24:2 | 11.1 ± 1.6 | 3.14 ± 0.8 | |
| 24:1 | 63.6 ± 10.5 | 22.2 ± 6.6 | |
| 24:1● | 32.6 ± 5.9 | 20.9 ± 2.4 | |
| 24:0 | 365 ± 85 | 285 ± 42 | |
| TOTAL | 729 ± 103 | 490 ± 103 ** | |
| Dihydro-ceramides (DH-Cer) | 16:0 | 13.6 ± 2.6 | 6.46 ± 2.3 |
| 18:0 | 2.98 ± 0.9 | 1.48 ± 1.4 | |
| 20:0 | 1.31 ± 0.2 | 0.78 ± 0.51 | |
| 22:0 | 12.0 ± 2.3 | 5.92 ± 2.5 | |
| 24:1 | 20.7 ± 11.4 | 12.4 ± 9.6 | |
| 24:0 | 60.8 ± 12.5 | 48.4 ± 28.5 | |
| TOTAL | 123 ± 32 | 75.4 ± 42.9 * | |
| Sphingomyelines (SM) | 14:1 | 4.15 ± 0.4 | 2.15 ± 0.6 |
| 14:0 | 84.0 ± 25.7 | 55.9 ± 12.3 | |
| 16:1 | 89.3 ± 17.5 | 68.0 ± 8.6 | |
| 16:0 | 1783 ± 662 | 1776 ± 477 | |
| 18:1 | 38.6 ± 5.8 | 22.7 ± 3.8 | |
| 18:0● | 140 ± 44 | 107 ± 16 | |
| 20:1 | 7.59 ±2.2 | 3.65 ± 1.5 | |
| 20:0 | 34.3 ± 6.9 | 24.6 ± 9.9 | |
| 22:2 | 3.57 ± 0.4 | 1.56 ± 0.4 | |
| 22:1 | 35.8 ± 2.5 | 21.3 ± 7.5 | |
| 22:1● | 47.7 ± 7.0 | 26.9 ± 4.9 | |
| 22:0 | 170 ± 24 | 125 ± 32 | |
| 24:3 | 6.83 ± 1.5 | 2.85 ± 1.1 | |
| 24:2 | 67.6 ± 24.3 | 31.1 ± 12.2 | |
| 24:1 | 437 ± 122 | 195 ± 25 | |
| 24:1● | 86.7 ± 22.5 | 72.1 ± 29.3 | |
| 24:0 | 665 ± 162 | 562 ± 71 | |
| TOTAL | 3555 ± 432 | 3139 ± 474 | |
| Dihydro-sphingomyelines (DH-SM) | 14:0 | 11.6 ± 2.9 | 3.26 ± 0.7 |
| 16:0 | 315 ± 104 | 76.8 ± 10.0 | |
| 18:0 | 26.7 ± 6.1 | 6.69 ± 2.6 | |
| 20:0 | 11.1 ± 2.9 | 3.87 ± 2.0 | |
| 22:0 | 50.2 ± 10.2 | 13.0 ± 4.8 | |
| 24:1 | 131 ± 46 | 23.9 ± 9.3 | |
| 24:0 | 201 ± 22 | 52.9 ± 14 | |
| TOTAL | 728 ± 72 | 166 ± 23 *** | |
| Glucosyl-ceramides (GlcCer) | 14:0 | 2.88 ± 0.5 | 2.80 ± 0.7 |
| 16:0 | 71.4 ± 4.4 | 100 ± 8 | |
| 18:0 | 8.67 ± 2.1 | 8.45 ± 2.1 | |
| 20:0 | 2.21 ± 0.8 | 1.78 ± 0.8 | |
| 22:0 | 27.3 ± 8.3 | 30.9 ± 8.2 | |
| 24:1 | 18.5 ± 7.3 | 13.7 ± 5.2 | |
| 24:0 | 146 ± 27 | 281 ± 49 | |
| TOTAL | 284 ± 41 | 429 ± 67 ** | |
| Glucosyl-dihydro-ceramides (GlcDH-Cer) | 16:0 | 17.7 ± 4.7 | 8.4 ± 2.6 |
| 18:0 | 1.78 ± 0.4 | 0.66 ± 0.43 | |
| 20:0 | 1.45 ± 0.1 | 0.66 ± 0.43 | |
| 22:0 | 12.4 ± 1.4 | 4.63 ± 1.3 | |
| 24:0 | 56.5 ± 10.4 | 38.5 ± 7.6 | |
| TOTAL | 87.9 ± 10.4 | 57.1 ± 16.3 ** | |
| Lactosyl-ceramides (LacCer) | 16:0 | 214 ± 19 | 33.5 ± 4.5 |
| 18.0 | 23.3 ± 3.9 | 1.94 ± 0.5 | |
| 20:0 | 6.81 ± 0.7 | 0.65 ± 0.45 | |
| 22:0 | 53.6 ± 23.0 | 8.38 ± 5.5 | |
| 24:1 | 51.1 ± 23.0 | 3.74 ± 0.9 | |
| 24:0 | 224 ± 36 | 57.2 ± 18.0 | |
| TOTAL | 569 ± 64 | 105 ± 29 *** | |
| Lactosyl-dihydro-ceramides (LacDH-Cer) | 16:0 | 46.1 ± 10.0 | 1.31 ± 0.5 |
| 18:0 | 5.32 ± 0.9 | - | |
| 20:0 | 2.47 ± 1.3 | - | |
| 22:0 | 25.7 ± 11.7 | 0.97 ± 0.77 | |
| 24:1 | 23.7 ± 11.7 | 3.49 ± 4.2 | |
| 24:0 | 91.8 ± 43.3 | 8.64 ± 4.3 | |
| TOTAL | 195 ± 60 | 14.4 ± 8.9 *** |
Values are expressed as pmol/106 cells (mean±SD; n=7; p-values are adjusted using a Holm method, *p<0.05, **p<0.01, ***p<0.001). To calculate the totals ± standard deviations per lipid species, we first calculated per repeat the sums of all chain length values, and then calculated the average and standard deviation of these sums. Due to the fact that in this high-throughput analysis, a peak was not present for each chain length in every repeat, some totals listed do not equal the mathematical sum of the individual chain length values.
R, total number of carbons:total number of insaturated acyl moieties.
●
Some species are detected with two independent peaks in the MS.