Fig 2: Increased microglial activation at Aß plaques can be observed in the offspring generated by the transgenic mother following ACI-24 vaccination. Statistical analysis of the average number of microglia recruited at the Aß plaques (plaque-associated microglia) of 5 control and 6 ACI-24 vaccinated mice (transgenic mother offspring) (A). The number of Iba1+ plaque associated microglia was normalised to the Aß area (as stained with 3552 antibody). Average CD68 signal was measured and normalised to the microglia number, revealing a trend towards increased CD68 coverage at the Aß plaques (B). Double staining of Iba1 and NLRP3 reveals an increase in the proportion of NLRP3+ microglia at the Aß plaque (as stained with Methoxy-X04) following ACI-24 vaccination (C,D). Scale bar: 20 µm. Graphs are presented as mean ± SEM (n.s. non-significant, * p < 0.05, unpaired two-tailed Student’s t-test).

Fig 3: Plaque-distant microglia show increased ramification following ACI-24 vaccination. Representative images and the respective Sholl analysis of the plaque-distant microglia (8–14 microglia per brain) in 5 control (in total 48 microglia) and 6 ACI-24 (in total 63 microglia) treated mice (transgenic mother offspring) (A). Microglia are depicted with Iba1, Aß is visualised using 3552 antibody and nuclei using Hoechst. Scale bar: 50 µm. Statistical analysis representing graphs for the ramification index (B), circularity (C), area (D) and perimeter (E) of Iba1+ cells. Graphs are presented as mean ± SEM (n.s. non-significant, ** p < 0.001). Kolmogorov–Smirnov test was performed to check if samples are following normal distribution, and then an unpaired two-tailed Student’s t-test or a Mann–Whitney test were carried out accordingly.