Dyn1 knockdown increases cell surface BACE-1 and holoAPP levels.

<p>A) Dyn KD fibroblast cells were treated with tamoxifen to induce dyn1 knockdown and then labeled with biotin followed by immunoprecipitation with streptavidin beads to pull down surface BACE-1. The amount of endogenous BACE-1 at cell surface was compared between control and dyn1 KD conditions. Total protein levels of endogenous BACE-1 were also determined from lysates as input. Protein levels were normalized to β-actin content and expressed as percentage of control. Data were collected in duplicate or triplicate from three independent experiments. Elevations in BACE-1 at cell surface (*<i>p<</i>0.05) without significant changes in total protein levels, were observed upon dyn1 KD. Cell lysates of BACE-1 expressing 293 cells were used as positive controls in these experiments. B) Alternatively, the amount of surface holoAPP was compared between control and DKO conditions. Elevations in holoAPP at cell surface (*<i>p<</i>0.05) without significant changes in total protein levels, were observed upon dyn1 KD. The sAPPα levels were determined in N2a 695 cells after dyn1 siRNA transfection, by immunoprecipitation with MAB348 followed by immunoblotting with 6E10 antibody. The levels of sAPPα was increased by 286.3% (<i>p</i> = 0.042) upon dyn1 KD.