Fig. 6.

TR is associated with AβPP and affects the level of sAβPPα. A) N2a-AβPP cells were mock-transfected (Mock), transfected with TR-V5/His or with LacZ-V5/His. The recombinant proteins and associated proteins in the cell extracts were then pulled down by anti-His beads. Western blot (WB) using 6E10 showed that AβPP was pulled down together with recombinant TR. B) Extracts of N2a cells mock transfected, transfected with TR-V5/His or LacZ-V5/His were incubated with GST-AβPP. The recombinant GST-AβPP and associated proteins were then pulled down by GS4B agarose beads. WB using anti-V5 showed that two TR-related proteins were pulled down together with AβPP, which correspond to the full-length (FL-TR) and a C-terminal fragment (CTF-TR) of TR. C) N2a-AβPP cells were mock-transfected, transfected with a TR-expressing plasmid, a control siRNA, or a siRNA specific to TR to downregulate TR. The level of sAβPPα in the 24-h conditioned medium was evaluated by WB using an antibody specific to sAβPPα. The level of TR transcript was estimated by RT-PCR. D) The bar graph shows the band intensity of sAβPPα in each condition adjusted by the respective band intensity of β-actin and then normalized to that of mock transfection (Mock) in the WB. n = 3, *p < 0.05 and **p < 0.001.