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. Author manuscript; available in PMC: 2015 May 13.
Published in final edited form as: Nature. 2014 Oct 12;515(7526):274–278. doi: 10.1038/nature13800

Figure 2. Robust increases of extracellular Aβ deposits in 3D-differentiated hNPCs with FAD mutations.

Figure 2

a. Thin layer 3D culture protocols (IF, immunofluorescence; HC, histochemical; IHC, immunohistochemical staining). b. Aβ deposits in 6-week differentiated control and FAD ReN cells in 3D Matrigel (green, GFP; blue, 3D6; scale bar, 25 μm; arrowheads, extracellular Aβ deposits; right-most panels, 3D6 staining was pseudo-colored to red). c. Select confocal z-stack images of 3D6-positive Aβ deposits. Z-sections with an interval of 2 μm were captured and the sections #1,3–4, #6 and #19 are shown (green, GFP; red, 3D6). d. IHC of Aβ deposits in ReN-mGAP cells. 3D-differentiated cells were treated with 1 μM BACE inhibitor IV, 500 nM DAPT, 500 nM SGSM41 or DMSO (brown, DAB (BA27); blue, hematoxylin; scale bar, 25 μm; arrowheads, large Aβ deposits). e. Detection of amyloid plaques with Amylo-Glo, a fluorescent amyloid-specific dye (Green, GFP; blue, Amylo-Glo; arrows, Amylo-Glo positive aggregates).