FIGURE 7.
LR11 Ser-2206 is necessary for LR11-mediated Aβ reduction. A, HEK293 cells were transfected with empty vector (Vector ctl), V5-LR11, orV5-LR11S2206A and harvested after 72 h. Media were conditioned for 48 h beginning 24 h after transfection. Equivalent volumes of conditioned media (Media) and 50 μg of protein for cell lysates (Lysate) were loaded for SDS-PAGE. Immunoblot analyses indicate similar levels of secreted APPsα and full-length APP among all samples. Calnexin was used as a loading control. B, endogenous secreted Aβ1–40 was detected by sandwich ELISA. Expression of V5-LR11 reduced secreted Aβ1–40 by 44% while expression of LR11S2206A reduced secreted Aβ1–40 by only 28% compared with empty vector control. Data shown are representative of three independent experiments. wt, wild-type.