(A) Confocal micrographs of GUVs incubated 24 h with FITC-tagged wildtype or mutant (tau-MBD or tau-C291/322A) tau peptides (20 µM) and Alexa647 (2 μM). Rhodamine-PE was incorporated into GUV preparation as a membrane marker. Scale bar 5 µM. Average profile plots for FITC-tau (B) and Alexa647 (C) for all tau peptides in addition to average Rhodamine-PE values for wild type peptide to indicate the position of the GUV membrane. (D,E) Quantitative analysis of membrane translocation (D) and pore formation (E) of wildtype and mutant tau (tau-MBD or tau-C291/322A) peptides. Data were derived from the same experimental GUVs as in panels (B,C). Data are presented as mean ± SEM. *P<0.05, **P<0.01, ****P<0.0001 (Welch’s ANOVA). For each tau peptide 95–99 individual GUVs were analysed. (F,G) Membrane translocation (F) and pore formation (G) of wildtype tau, DTT-treated wildtype tau and tau-C291/322A peptides diluted in PBS and preincubated 1 h with or without DTT before addition of GUVs and Alexa647 and final 24-h incubation. Data are presented as mean ± SEM. *P<0.05, ****P<0.0001 (Welch’s ANOVA). For each tau peptide condition 103–112 individual GUVs were analysed.