Figure 5

SNX12 interacts with BACE1. (A) HeLa cells were co-transfected with SNX12-EGFP (in green) and BACE1-HA. Cells were then fixed, permeabilized and immunostained sequentially with primary antibodies against HA and secondary antibodies conjugated with Alexa Fluor 594 (to indicate BACE1, in red). Samples were examined by a fluorescence confocal microscope. (B) BACE1-HA was co-transfected with control (−) or SNX12-Myc (+) into HEK 293T cells. Equal protein amounts of cell lysates were used for immunoprecipitation (IP) with an anti-HA antibody or control IgG. Immunoprecipitated proteins were subjected to Western blot with anti-HA or anti-Myc antibodies to detect BACE1 or SNX12, respectively. Ten percent of cell lysates were used as input. (C) HEK-APP_Swe cells were transfected with control (−) or SNX12-Myc (+). Equal protein amounts of cell lysates were subjected to IP with control IgG or the anti-APP antibody 369. Immunoprecipitated proteins and input were subjected to Western blot. (D) Equal protein amounts of SH-SY5Y cell lysates were subjected to IP with control IgG, anti-SNX12 antibody, or the anti-BACE1 antibody 689. Immunoprecipitated proteins and input were subjected to Western blot analysis.