Fig. 3.

Treatment with mDF2β reduces survival of XS52 cells (A and D) and bone marrow-derived DCs (BMDCs; B and C). Cells, treated for the indicated days with 5 μg/ml mDF2β-sFv38 (mDF2b), sFv315, or 10 ng/ml LPS, with or without 5 μg/ml polymixin B. To assess cell survival, cells were stained with Annexin V/propidium iodide (PI; A, B, and D). In parallel, cells were stained with antibodies for CD11c⁺/CD40⁺/CD86⁺ to assess cell activation (C). Inhibition of pan-caspases using ZVAD exaggerates the mDF2β-induced killing, leading to the cell death after only overnight incubation (D). XS52 cells were incubated overnight with 5 μg/ml mDF2β or sFv315 in the presence or absence of ZVAD-fmk or control ZFA-fmk. All results shown (A-D) were independently reproduced at least three times, and the mean of the representative triplicate experiment is shown ± sem. P values are for comparisons of pooled mDF2β data with LPS, mDF2β + LPS, or mDF2β + polymixin B (A) or between groups indicated by solid lines (D).