Reexpression of L-selectin ligand activity after neuraminidase or protease treatment of KG1a cells is dependent on de novo protein synthesis
| KG1a treatment3-150 . | Mean (SEM) % control binding3-151 . |
|---|---|
| Neuraminidase (t = 0, no culture) | 0.2 (0.1)+ |
| Neuraminidase buffer control (no culture) | 98.5 (7.5) |
| Neuraminidase, 24-h culture | 101.3 (6.5) |
| Bromelain (t = 0, no culture) | 3.8 (0.4)++ |
| Bromelain, 20-h culture | 95.2 (7.1) |
| Neuraminidase, 20-h culture with cycloheximide (1.25 μg/mL) | 0.5 (0.2)+ |
| Bromelain, 20-h culture with cycloheximide (1.25 μg/mL) | 0.6 (0.3)++ |
| 20-h culture with cycloheximide (1.25 μg/mL) (no neuraminidase or bromelain digestion) | 91.8 (5.8) |
| KG1a treatment3-150 . | Mean (SEM) % control binding3-151 . |
|---|---|
| Neuraminidase (t = 0, no culture) | 0.2 (0.1)+ |
| Neuraminidase buffer control (no culture) | 98.5 (7.5) |
| Neuraminidase, 24-h culture | 101.3 (6.5) |
| Bromelain (t = 0, no culture) | 3.8 (0.4)++ |
| Bromelain, 20-h culture | 95.2 (7.1) |
| Neuraminidase, 20-h culture with cycloheximide (1.25 μg/mL) | 0.5 (0.2)+ |
| Bromelain, 20-h culture with cycloheximide (1.25 μg/mL) | 0.6 (0.3)++ |
| 20-h culture with cycloheximide (1.25 μg/mL) (no neuraminidase or bromelain digestion) | 91.8 (5.8) |
After neuraminidase or bromelain treatment, all cells were washed before culturing as indicated.
Number of lymphocytes adherent to confluent area of KG1a were counted by light microscopy using an ocular grid under 100× magnification (quantified a minimum of 3 fields per slide, 3 slides per experiment, 3 separate experiments). Results are presented as percentage binding compared with corresponding untreated control KG1a cytospin preparations.
+,++Statistically significant (P < .01, by paired t test) difference compared with undigested, neuraminidase buffer-incubated (+) or RPMI 1640-incubated (++) cells. All other values are not statistically different (P > .05).