Anti cd95 fas pe cy7

Murine and human carcinoma cells were treated for 72 h with 10 μM ARI-4175 and assessed for changes in a variety of immunologically relevant cell-surface molecules [17 (link)]. Murine cells were stained using the following antibodies: anti-H2Kb/H2Db-FITC, anti-H2Kd/H2Dd-APC, anti-CD54 (ICAM-1)-PE, anti-CD95 (Fas)-PE-Cy7 (BD Biosciences, San Diego, CA), anti-Col-1 (CEA)-FITC ([18 (link)]), and anti-Calreticulin-PE (R&D Systems, Minneapolis, MN). Human cells were assessed with: anti-CD95 (Fas)-PerCP-Cy5.5, anti-CD54 (ICAM-1)-PE, anti-CD227 (MUC1)-FITC, anti-HLA-A2-FITC, (BD Biosciences), anti-CD66 (CEA)-APC (Miltenyi Biotech, Auburn, CA) and anti-Calreticulin-PE (R&D Systems). Stained cells were acquired on an LSR II flow cytometer and analyzed using FloJo software. Isotype staining was < 5% for all samples analyzed. Proteins were scored as up-regulated if either the percent of cells or the mean fluorescence intensity (MFI) was increased by > 10% relative to untreated and vehicle-treated controls.

Single-cell suspensions from the spleens and livers of CLT and CLT; AID^−/− RDBC chimeras and control mice were stained with each of the following sets of anti-mouse monoclonal antibodies: anti-B220-PE-Cy5 (eBioscience Inc.) and anti-IgM-FITC (eBioscience Inc.); anti-B220-APC (eBioscience Inc.) and anti-Igκ-PE (BD Biosciences); anti-CD19-FITC (BD Biosciences) and anti-CD95/Fas-PE-Cy7 (BD Biosciences); or anti-CD8a-APC (BD Biosciences) and anti-CD4-FITC (eBioscience Inc.). FACS data were acquired using the FACSCalibur Flow Cytometer equipped with CellQuest software (BD Biosciences) and analyzed with FlowJo software (TreeStar).