Figure 7.
Figure 7. Chemokine release depends on caspase-8 activity in the absence of apoptosis. (A) Cells were incubated for 4 hours with different amounts of cross-linked FasL, with or without cycloheximide (CHX) and apoptotic cells were scored by FACS using Annexin V staining; CD56-CD8+ cells (□) and CD56+CD8+ cells (▪). (B) CD56+CD3+ cells were pretreated for 1 hour with different inhibitors, washed, and used in indirect neutrophil migration assays. The horizontal line at migration index 1.0 indicates lack of chemotaxis. Concentrations used were 100 μM for the cell permeable caspase-1 inhibitor (CP YVAD-CHO) and the cell permeable caspase-8 inhibitor (CP IETD-CHO), 10 μg/mL for actinomycin D, 1 μL/mL GolgiPlug (brefeldin A), 200 μM for NDGA, and 300 μM for indomethacin. Data shown are the mean ± SEM of triplicates from 1 donor.

Chemokine release depends on caspase-8 activity in the absence of apoptosis. (A) Cells were incubated for 4 hours with different amounts of cross-linked FasL, with or without cycloheximide (CHX) and apoptotic cells were scored by FACS using Annexin V staining; CD56-CD8+ cells (□) and CD56+CD8+ cells (▪). (B) CD56+CD3+ cells were pretreated for 1 hour with different inhibitors, washed, and used in indirect neutrophil migration assays. The horizontal line at migration index 1.0 indicates lack of chemotaxis. Concentrations used were 100 μM for the cell permeable caspase-1 inhibitor (CP YVAD-CHO) and the cell permeable caspase-8 inhibitor (CP IETD-CHO), 10 μg/mL for actinomycin D, 1 μL/mL GolgiPlug (brefeldin A), 200 μM for NDGA, and 300 μM for indomethacin. Data shown are the mean ± SEM of triplicates from 1 donor.

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