Treatment of DC with CHX or Bis results in decreased levels of c-FLIPL and sensitivity to CD95-mediated apoptosis.
(A) DC were incubated in medium alone or in the presence of either CHX (10 μg/mL) or Bis III (20 μmol/L). At the indicated times, whole-cell extracts were electrophoresed and probed with anti-FLIPL mAb (NF6) followed by an appropriate second Ab. Data are representative results from 3 experiments done on samples from different blood donors. (B) DC were incubated with agonistic anti-CD95 mAb (0.5 μg/mL) in the presence of CHX (10 μg/mL) or Bis III (20 μmol/L). After 4, 8, or 24 hours, cells were fixed, DNA was stained with PI, and the percentage of hypodiploid nuclei (subG1 peak) was analyzed with use of flow cytometry. Data are representative results from 2 experiments done on samples from different blood donors.
