Fig 2: Semi-quantitative analysis of immunohistochemical data of proteins with altered expression in CAA type-1. Immunohistochemical stainings were quantified by measuring the percentage of pixels that showed positive immunoreactivity. Significance was calculated using a one-way ANOVA (Kruskal-Wallis test) and posthoc Dunn’s Multiple Comparison Test. A significant increase (p < 0.05) in immunoreactivity in the CAA group compared to both control and AD groups was observed for NDP and COL6A2. For APOE, APCS and HTRA1 significant differences were only found when comparing CAA with control, but not with the AD group. Data are expressed as mean ± SEM

Fig 3: Immunoblotting analysis of proteins with altered expression in CAA type-1. a Immunoblotting was performed for, NDP, COL6A2, APCS and APOE on occipital lobe tissue lysates of non-demented controls (N) AD cases (A) and CAA cases (C). Immunoreactivity was observed at the correct molecular weight for each protein. b A significant difference (p < 0.05) was found in the expression of NDP in the CAA group when compared to control but not when compared to the AD group. For COL6A2, APCS and APOE significance was not reached between any of the groups using this technique. Data are expressed as mean ± SEM

Fig 4: Expression of CAA type-1 markers in other small vessel diseases. IHC for Aß, NDP, COL6A2, APCS and APOE was performed. In a CAA type-1 case immunoreactivity for all marker proteins is confirmed (a-e). Also immunoreactivity is seen for all markers in the cotton wool case and Aß pathology was confirmed (f). For NDP, APOE and APCS immunoreactivity is also seen localizing to severe dyshoric angiopathy (g, i and j). COL6A2 immunoreactivity is restricted to the vessel wall (h). In de Prp-CAA case Aß pathology was absent (k). Extensive immunoreactivity was observed for NDP, COL6A2, APOE and APCS (l-o). No immunoreactivity was observed for Aß, NDP, COL6A2, APOE and APCS in the white matter of control tissue (p-t). In the CADASIL case no Aß pathology was present (u). Mild immunoreactivity for NDP (v) COL62A staining was most pronounced (w). APOE and APCS also displayed mild immunoreactivity related to the affected vessels (x, y). In hypertension related small vessel disease no Aß was detected (z) immunoreactivity of COLA6A2 was moderate (ab) while immunoreactivity for NDP, APOE and APCS were low but present (aa, ac and ad). In the CARASAL case only prominent immunoreactivity of COL6A2 was seen in affected vessels (ag) while NDP, APOE and APCS were absent (af, ah and ai). Scale bar in (a) indicates 100 µm