Effect of TGF-β1 on Fas Expression on Lin− Murine BM Cells
| Cytokines . | Specific Mean Fluorescence Intensity . | Mean Fluorescence, . | ||
|---|---|---|---|---|
| . | Exp 1 . | Exp 2 . | Exp 3 . | % of TGF-β1–Untreated Control (mean of 3 experiments) . |
| None (freshly isolated cells) | 4.2 | 2.8 | 5.8 | NA |
| GM-CSF | 8.6 | 8.5 | 23.9 | 100 |
| GM-CSF + TGF-β1 200 ng/mL | 6.2 | 3.9 | 13.2 | 57 |
| GM-CSF + TGF-β1 20 ng/mL | 6.6 | 4.1 | 12.2 | 56 |
| GM-CSF + TGF-β1 2 ng/mL | 7.5 | 2.5 | 13.1 | 56 |
| GM-CSF + TGF-β1 0.2 ng/mL | 8.5 | 5.9 | 17.6 | 78 |
| GM-CSF + TGF-β1 0.02 ng/mL | 10.5 | 8.6 | ND | 112* |
| GM-CSF + TNF | 13.8 | 13.0 | 37.3 | 100 |
| GM-CSF + TNF + TGF-β1 200 ng/mL | 8.9 | 6.0 | 19.5 | 54 |
| GM-CSF + TNF + TGF-β1 20 ng/mL | 7.3 | 3.6 | 19.1 | 47 |
| GM-CSF + TNF + TGF-β1 2 ng/mL | 10.1 | 5.4 | 23.5 | 61 |
| GM-CSF + TNF + TGF-β1 0.2 ng/mL | 13.8 | 7.3 | 23.6 | 70 |
| GM-CSF + TNF + TGF-β1 0.02 ng/mL | ND | 8.7 | 28.5 | 74* |
| Cytokines . | Specific Mean Fluorescence Intensity . | Mean Fluorescence, . | ||
|---|---|---|---|---|
| . | Exp 1 . | Exp 2 . | Exp 3 . | % of TGF-β1–Untreated Control (mean of 3 experiments) . |
| None (freshly isolated cells) | 4.2 | 2.8 | 5.8 | NA |
| GM-CSF | 8.6 | 8.5 | 23.9 | 100 |
| GM-CSF + TGF-β1 200 ng/mL | 6.2 | 3.9 | 13.2 | 57 |
| GM-CSF + TGF-β1 20 ng/mL | 6.6 | 4.1 | 12.2 | 56 |
| GM-CSF + TGF-β1 2 ng/mL | 7.5 | 2.5 | 13.1 | 56 |
| GM-CSF + TGF-β1 0.2 ng/mL | 8.5 | 5.9 | 17.6 | 78 |
| GM-CSF + TGF-β1 0.02 ng/mL | 10.5 | 8.6 | ND | 112* |
| GM-CSF + TNF | 13.8 | 13.0 | 37.3 | 100 |
| GM-CSF + TNF + TGF-β1 200 ng/mL | 8.9 | 6.0 | 19.5 | 54 |
| GM-CSF + TNF + TGF-β1 20 ng/mL | 7.3 | 3.6 | 19.1 | 47 |
| GM-CSF + TNF + TGF-β1 2 ng/mL | 10.1 | 5.4 | 23.5 | 61 |
| GM-CSF + TNF + TGF-β1 0.2 ng/mL | 13.8 | 7.3 | 23.6 | 70 |
| GM-CSF + TNF + TGF-β1 0.02 ng/mL | ND | 8.7 | 28.5 | 74* |
50,000 Lin− BM cells were cultured for 44 hours in 96-well microtiter plates in complete IMDM in the presence of GM-CSF (20 ng/mL), in the absence or presence of TNF-α (20 ng/mL) and increasing concentrations of TGF-β1 , as indicated. Freshly isolated cells and groups of cultured Lin− cells were stained with an FITC-conjugated anti-Fas antibody (Jo2) or FITC-conjugated irrelevant control hamster IgG. Samples were analyzed by flow cytometry as described in Materials and Methods. For each of three individual experiments, results are displayed as mean specific fluorescence intensity (relative mean fluorescence of control antibody is subtracted from fluorescence obtained with Jo2 antibody). In addition, the fluorescence (mean of three experiments) is expressed as percentage of TGF-β1–untreated control cells. The mean fluorescence intensity of control antibody for various samples varied between 2.5 and 5.7.
Abbreviations: NA, not applicable; ND, not determined.
Mean of two experiments.