Fig. 5: FORTIS can detect endocytosis induced by Aβ.

a Hippocampal neurons overproducing Aβ42 due to the expression of APP_swe/lnd. The medium of these cultures that contains Aβ42 was used to induce AMPAR endocytosis in c and d. b A western blot of lysates of neurons expressing either EGFP or APP_swe/lnd and probed with an anti-β-Amyloid antibody only recognizes an APP and Aβ band in the cultures expressing APP_swe/lnd. c A Heat map of SEP-GluA1 and pHuji-GluA1 fluorescence where each square represents a single cortical culture in a 96-well plate following treatment with medium taken from neurons expressing APP_swe/lnd and secreting Aβ42 (see a). d Quantification of the experiment shown in c, showing the individual relative fluorescence (each circle represents a single culture) following treatment with the Aβ42-containing medium. Statistical significance was calculated according to the Mann–Whitney test. e Heat map of SEP-GluA1 relative fluorescence where each square represents a single hippocampal culture in a 96-well plate following treatment with synthetic Aβ42 (3 or 4 μM), with or without preincubation with the PTEN-PDZ peptide (5 or 10 μM). f Quantification of the experiment shown in e, showing the individual relative fluorescence (each circle represents a single culture) following treatment with synthetic Aβ42 and the PTEN-PDZ peptide. Statistical significance was calculated according to the Mann–Whitney test. g cLTP was induced in cultures treated with Aβ42 (4 μM), with or without preincubation with the PTEN-PDZ peptide (10 μM). Statistical significance was calculated according to two-way ANOVA followed by Tukey’s multiple comparisons post hoc tests, and the data are presented as the mean ± SEM.