Fig. 5.
Effect of endocytosis inhibition on induction of apoptosis by CVPE, CIVUV, or CIV. (A) Cells were pretreated with ammonium chloride (AC; 10 mM) for 30 min prior to addition of CVPE (10 μg/ml), CIVUV (10 μg/ml), or CIVXS (400 μg/ml) and incubated at 21 °C for 24 h. (B) Cells were pretreated with bafilomycin A1 (Bf; 1 μM) for 30 min prior to addition of CVPE (10μg/ml), CIVUV (10μg/ml), or CIV (400 μg/ml) and incubated at 21 °C for 24 h. (C) CIVUV (10μg/ml) was attached to polysterene beads as described in Materials and methods. Cells were challenged with this complex, incubated at 21 °C for 24 h, and stained with safranin to differentiate between blebs and beads. Bars in plates A, B, and C represent percent blebbing based on counting 200 cells per field. Error bars indicate standard deviations for four independent experiments per set.