Figure 6.

FAIM-L is necessary for TNFα protection against Aβ-induced death. (a) Neurons were infected with shFAIM-L or the corresponding scrambled (sc) control for 6 days and then treated for 48 h with 5 μM ADDLs and 100 ng/ml TNFα. Counts of nuclei after Hoechst staining were used to determine cell death. (b) Primary neurons treated as in (a) were processed for western blot analysis of caspase-3 cleavage and FAIM-L expression. Pan-ERK was used as a loading control. (c) Scheme showing how the lentivirus-expressing silent mutated FAIM-L (smFAIM-L) avoids shRNA downregulation, giving the expression of FAIM-L. (d) Expression of FAIM-L after 6 days of infection with the indicated lentiviruses; *film least exposed shows that when both lentiviruses are expressed (shFAIM-L+smFAIM-L), the shFAIM-L downregulates endogenous FAIM-L. Loading control was determined with pan-ERK. (e) Neurons were infected as indicated and treated for 48 h with 5 μM ADDLs and 100 ng/ml TNFα. Cell death was determined by counting nuclei after Hoechst staining. In all cases, data are mean±S.E.M. of three independent experiment, except for (b) that is six. One-way ANOVA with Newman–Keuls test. ***P<0.001, **P<0.01 and *P<0.05 for the same treatment; ^##P<0.01 and ^#P<0.05 between ADDLs and ADDLs+TNFα