PTX3 recognition requires cell death and does not depend on the stimulus triggering apoptosis
| Cells . | Treatment . | Relative fluorescence intensity . |
|---|---|---|
| Alive | none | 1 |
| Permeabilized | saponin | 1.2 |
| Apoptotic | cycloheximide | 10 |
| staurosporin | 19.8 | |
| PDTC | 12.3 | |
| BAPTA | 11.7 | |
| dexamethasone | 17.9 | |
| UV irradiation (16 h) | 22.4 | |
| Postapoptotic | UV irradiation (48 h) | 2.8 |
| Necrotic | freezing | 7.3 |
| boiling | 2.4 | |
| hyperosmotic shock | 5.6 |
| Cells . | Treatment . | Relative fluorescence intensity . |
|---|---|---|
| Alive | none | 1 |
| Permeabilized | saponin | 1.2 |
| Apoptotic | cycloheximide | 10 |
| staurosporin | 19.8 | |
| PDTC | 12.3 | |
| BAPTA | 11.7 | |
| dexamethasone | 17.9 | |
| UV irradiation (16 h) | 22.4 | |
| Postapoptotic | UV irradiation (48 h) | 2.8 |
| Necrotic | freezing | 7.3 |
| boiling | 2.4 | |
| hyperosmotic shock | 5.6 |
Jurkat cells untreated, permeabilized, and killed by apoptosis or necrosis were challenged with biotinylated PTX3 (10 μg/mL) and fluorescein isothiocyanate (FITC)–streptavidin. The binding was then assessed by flow cytometry, as described in “Materials and methods.” Results are expressed as relative fluorescence intensity calculated by dividing the mean fluorescence intensity in the presence of PTX3 and FITC-streptavidin and in the presence of FITC-streptavidin alone. Results are representative of 4 independent experiments.
PDTC indicates pyrrolidine dithiocarbamate; BAPTA, 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid tetra(acetoxymethyl) ester.