Figure 1.

NF-κB and caspases show parallel activation. (A–C) HeLa-CD95 cells were stimulated with (A)1500 ng/ml, (B) 500 ng/ml and (C) 250 ng/ml of agonistic anti-CD95 antibodies for the indicated periods of time. The cellular lysates were analyzed by western blotting using antibodies against caspases, IκBα and p-IκBα. Nonspecific bands of anti-p-IκBα are marked (NS). Results are representative of four different experiments. (D) To follow p65 localization, Hela-CD95 cells were stably transfected with p65–mCherry. Cells were stimulated with 1500 ng/ml anti-CD95 antibody and imaged using fluorescent microscopy over the respective time. Measurements of mCherry (depicted in gray) indicated translocation of p65 on CD95 activation. Induced apoptosis is observed by the appearance of apoptotic bodies. Scale bar: 10 μm.