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Fig. 2. Arnt-null TS cell differentiation can lead to granzyme B-mediated apoptosis. (A) DAPI-staining of differentiated wild-type (upper panel) and mutant (lower panel) TS cells revealed a significant number of pyknotic nuclei indicative of apoptosis only in Arnt-null cells (arrows, lower panel). This impression was confirmed by FITC labeling of nicked DNA (inset). (B) Time-lapse video microscopy of Arnt-null cells was performed starting 4 days after differentiation. The results showed wide-scale breakdown of the plasma membrane and cytoplasmic vacuolization (arrows) that culminated in cell sloughing from the tissue culture plate (data not shown). (C) Cytoplasmic (Cyt) and nuclear (Nuc) extracts of undifferentiated (U) and differentiated (D) wild-type and Arnt-null TS cells were subjected to immunoblot analysis using antibodies specific for granzyme B (GR B), poly (ADP-ribose) polymerase (PARP), tankyrase 2 (TANK2) and {alpha}–tubulin ({alpha}-TUB).