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. 2016 Dec 5;114(4):E476–E485. doi: 10.1073/pnas.1618657114

Fig. 4.

Fig. 4.

The Aβ42/Aβ40 ratios generated in liposome- and cell-based cleavage assays are similar to those obtained in detergent micelle-based assays. (A) A schematic diagram of the liposome-based cleavage assay. The WT and mutant γ-secretases were individually reconstituted into the liposomes together with APP-C99 and examined for their abilities to generate Aβ42 and Aβ40 using the AlphaLISA assay. (B) The Aβ42/Aβ40 ratios generated in the liposome-based cleavage assays are similar to those obtained in detergent micelle-based assays. Results from 11 representative γ-secretase variants are shown here. (C) A schematic diagram of the cell-based cleavage assay. The CRISPR/Cas9 technology was used to generate PS1−/− N2a cells. The pCAG vectors, each encoding WT or a distinct γ-secretase variant, were individually transfected into N2a cells for assessment of production of Aβ42 and Aβ40. (D) The Aβ42/Aβ40 ratios generated in the cell-based cleavage assays are similar to those obtained in detergent micelle-based assays.