Fig. 5

Characterization of TNFa-induced apoptosis in Smg7 −/−, Ces2a −/− and control cell lines. a The NF-κB responsive p65 protein was detected by high-content microscopy and the ratio of nuclear:cytosolic forms was compared for the mutant cell lines in untreated and TNFa treated conditions. b The NF-κB transcriptional response was measured by luciferase assay and fold induction calculated relative to a transfection control. c Smg7 −/−, Ces2a −/−, Hnrnpf −/− cell lines can be resensitized to TNFa by addition of cycloheximide (CHX) but are significantly less sensitive than the control. Trnfrsf1a −/− cells cannot be resensitized. d Cell death proteins were detected by Western blot at 0 and 16 h of TNFa treatment. Basal levels of p53 are upregulated in Ces2a −/− and in Smg7 −/− cells following TNFa addition. e, f Control cells show a significant increase in nuclear (DAPI-stained) fraction of p53 (red) following TNFa treatment while Smg7 −/− cells show similar levels. *P < .05. **P < .01. ***P < .001. n.s. not significant. Scale bar is 50 μm