Figure 9.

Bad is required for p75NTR-induced apoptosis. PC12 cells were transfected with GFP plasmid alone or with GFP plasmid together with plasmids encoding DN-Bad (S128A) or expressing Bad-RNAi. Cells were infected 48 hr later with LacZ or p75NTR adenovirus and, at 24 hr after infection, were fixed and immunostained for cleaved caspase 3 as described in Materials and Methods. Control experiments established that cleaved caspase 3 immunoreactivity correlates with TUNEL staining and is thus a valid surrogate for direct measurement of apoptosis (see supplementary Fig. 1, available at www.jneurosci.org). Transfected cells were scored for caspase 3 cleavage by a blind observer (n = 300 cells/condition). ^**Indicates a difference of p < 0.001 between GFP/Mock (Bar 1) and GFP/p75NTR (Bar 5), and ^*indicates a difference of p < 0.001 between GFP/p75NTR (Bar 5) and both DN-Bad/p75NTR (Bar 6) and with Bad RNAi/p75NTR (Bar 7), indicated by ANOVA.