Supplementary Figure 6: OptiCAB assay in HEK293T cell lines.
From: Visualizing APP and BACE-1 approximation in neurons yields insight into the amyloidogenic pathway
(a) HEK293T cells were transfected with indicated constructs and visualized 5-6 h post-transfection. Note in the APP:VN/BACE-1:VC group, complemented APP/BACE-1 fluorescence was widespread and punctate (zoomed inset). However, diminished fluorescence was seen in in APP:VN/BACE-1:VC transfected cells treated with an inhibitor of clathrin-dependent endocytosis (Dynasore), or in cells transfected with APPYENPTY:VN/BACE-1:VC. Fluorescence complementation was also markedly attenuated when cells were transfected with APP(Ice):VN/BACE-1:VC.
(b)Western blot of HEK293T cells transfected with APP:VN/BACE-1:VC (in presence of a γ-secretase inhibitor) shows the expected ~30 kDa β-cleavage product of APP:VN (data from 2-3 independent cultures).
