Bcl-2 and Bcl-X expression in migrated and nonmigrated cells
| Cells . | Bcl-2* . | Bcl-X* . | PECAM-1* . |
|---|---|---|---|
| Before migration (CD14+) | 20 ± 10 | 18 ± 9 | ND |
| Migrated (CD14+CD34+) | 180 ± 21 | 160 ± 13 | 210 ± 20 |
| Nonmigrated (CD14+CD34−) | 66 ± 11 | 60 ± 8 | 35 ± 5 |
| Cells . | Bcl-2* . | Bcl-X* . | PECAM-1* . |
|---|---|---|---|
| Before migration (CD14+) | 20 ± 10 | 18 ± 9 | ND |
| Migrated (CD14+CD34+) | 180 ± 21 | 160 ± 13 | 210 ± 20 |
| Nonmigrated (CD14+CD34−) | 66 ± 11 | 60 ± 8 | 35 ± 5 |
Bcl-2 and Bcl-X expression was evaluated by cytoplasmic immunofluorescence and PECAM-1 expression by surface phenotype in unfractionated CD14+ cells or in cells migrated or not through HUVEC monolayers, collected after an additional 36 hours of culture. Cells were fixed in 2% paraformaldehyde, and for cytoplasmic fluorescence permeabilized with 0.01% NP-40, stained with the specific anti–Bcl-2, anti–Bcl-X, or anti–PECAM-1 mAbs followed by FITC-GAM, and run on a FACSort. At least 104 cells/sample were analyzed and results plotted as arbitrary units (au) of mean fluorescence intensity (MFI; mean ± SD from 8 experiments). MFI of cells stained with an FITC-irrelevant mAb was 15 ± 5 au. ND indicates not determined.