Figure 5.

Characterization of NP-vFLIP-α2-induced cell death in chronically infected macrophages. (a) After a single treatment with NP-vFLIP-α2 for 8 h, chronically infected macrophages were cultured for an additional 6 days. Cell lysates were assessed for cleaved CASP3 by western blot and analyzed using ImageJ software. (b) Cleaved CASP8 was also measured in the same cell lysates. Staurosporine 1 µM treatment for 6 h was used as a positive control. (c) Chronically infected macrophages were pretreated with 1 mM 3-methyladenine, 20 µM Z-VAD-FMK, 50 μM necrostatin-1 or 200 nM BAF for 2 h, followed by treatment with 10 µM NP-vFLIP-α2 for an additional 24 h. Cell culture supernatants were tested for cytotoxicity. Data are derived from 4 different donors and reported as means ± s.e.m. Representative immunoblots are shown. * P < 0.05, ** P < 0.01, *** P < 0.001.