Figure EV1. NCT ectodomain regulates the Aβ 42/40 ratio.

1. Representative SDS–PAGE/Western blot analysis of CHAPSO‐solubilized membrane proteins from Ncstn ^−/− or Psen1 ^−/− /Psen2 ^−/− MEF cell lines stably expressing WT or mutant mouse NCT/human PSEN1 subunits (shown in Fig 2A). The presence of mature, glycosylated NCT, N‐terminal, and C‐terminal fragments of the endoproteolyzed PSEN1 and PEN‐2, compared to NCT knock‐out (KO) or PSEN1/2 double KO (dKO) cells, indicates that WT and mutants reconstitute GSEC complexes. Arrowheads indicate the position of molecular weight markers.
2. Aβ peptides present in the extracellular media of cultured WT or mutant mNCT/hPSEN1 MEF cell lines, transiently expressing APP_C99 substrate, were quantified by ELISA, and the Aβ 42/40 ratio were determined and plotted as % of the WT condition. The data are represented as mean ± SD, N ≥ 4 independent experiments. One‐way ANOVA and Dunnett's post hoc test were used to determine the statistical significance; ***P > 0.001 and ****P > 0.0001 (compared to WT condition). F(DFn, DFd): F(31, 290) = 873,6.

Source data are available online for this figure.