Distribution of LacCer, Lyn, and marker enzymes in subcellular fractions of neutrophils
| Fraction . | MPO . | Lactoferrin . | Gelatinase . | ALP . | LDH . | LacCer . | Lyn . |
|---|---|---|---|---|---|---|---|
| Cytosol | 0.7 ± 0.2 | 0.0 | 2.7 ± 2.3 | 1.0 ± 0.3 | 95.4 ± 3.5 | 0.9 ± 0.2 | 0.0 |
| Plasma membrane | 1.5 ± 0.3 | 0.0 | 5.5 ± 1.3 | 55.8 ± 3.2 | 0.0 | 10.0 ± 0.9 | 99.5 ± 2.1 |
| Gelatinase glanules | 5.2 ± 1.2 | 7.9 ± 2.2 | 47.3 ± 1.2 | 15.4 ± 1.7 | 0.0 | 15.5 ± 2.1 | 0.0 |
| Specific granules | 11.6 ± 2.4 | 89.9 ± 5.1 | 42.3 ± 2.3 | 7.9 ± 0.8 | 0.0 | 36.5 ± 2.7 | 0.0 |
| Azurophil granules | 68.2 ± 4.2 | 0.2 ± 0.1 | 4.3 ± 2.4 | 10.3 ± 1.4 | 0.0 | 37.1 ± 3.2 | 0.0 |
| Recovery | 87.2 ± 5.1 | 98.0 ± 3.2 | 103.5 ± 4.1 | 90.3 ± 3.1 | 95.4 ± 3.5 | 99.0 ± 4.1 | 99.5 ± 2.1 |
| Fraction . | MPO . | Lactoferrin . | Gelatinase . | ALP . | LDH . | LacCer . | Lyn . |
|---|---|---|---|---|---|---|---|
| Cytosol | 0.7 ± 0.2 | 0.0 | 2.7 ± 2.3 | 1.0 ± 0.3 | 95.4 ± 3.5 | 0.9 ± 0.2 | 0.0 |
| Plasma membrane | 1.5 ± 0.3 | 0.0 | 5.5 ± 1.3 | 55.8 ± 3.2 | 0.0 | 10.0 ± 0.9 | 99.5 ± 2.1 |
| Gelatinase glanules | 5.2 ± 1.2 | 7.9 ± 2.2 | 47.3 ± 1.2 | 15.4 ± 1.7 | 0.0 | 15.5 ± 2.1 | 0.0 |
| Specific granules | 11.6 ± 2.4 | 89.9 ± 5.1 | 42.3 ± 2.3 | 7.9 ± 0.8 | 0.0 | 36.5 ± 2.7 | 0.0 |
| Azurophil granules | 68.2 ± 4.2 | 0.2 ± 0.1 | 4.3 ± 2.4 | 10.3 ± 1.4 | 0.0 | 37.1 ± 3.2 | 0.0 |
| Recovery | 87.2 ± 5.1 | 98.0 ± 3.2 | 103.5 ± 4.1 | 90.3 ± 3.1 | 95.4 ± 3.5 | 99.0 ± 4.1 | 99.5 ± 2.1 |
Neutrophils were disrupted by nitrogen cavitation, and the postnuclear supernatant was applied on top of a 3-layer Percoll gradient and centrifuged. The resultant 4 visible bands (corresponding to the azurophil granule, the specific granule, the gelatinase granule, and the plasma membrane fractions) and the cytosol fraction were collected. The contents of LacCer, Lyn, and marker enzymes in each fraction were expressed as percentage of total lysates. Data represent the means ± SD of 3 experiments.
MPO indicates myeloperoxidase; ALP, alkaline phosphatase; LDH, lactate dehydrogenase.