Fig. 3

The inflammatory activation (reactive astrocytes and microglia) in retinas from AD transgenic animals is dampened by 12A12mAb treatment. Retinal homogenates extracts from animals of three experimental groups (wild-type, Tg2576 and Tg2576 + mAb) were analyzed by Western blotting for inflammatory proteins (GFAP, Iba1) (a). Semi-quantitative densitometric analysis (n = 6) of intensity signals (b) indicates lower levels of GFAP and Iba1 in Tg2576 mice + mAb compared to not-immunized counterpart. β-actin was used as loading control. Arrows on the right side indicate the molecular weight (kDa) of bands calculated from migration of standard proteins. Values are from at least three independent experiments and statistically significant differences were calculated by one-way analysis of variance (ANOVA) followed by Bonferroni’s post-hoc test for multiple comparison among more than two groups. p < 0.05 was accepted as statistically significant (*p < 0.05; **p < 0.01; ***p < 0.0005; ****p < 0.0001). Representative images of retinal slices from animals of three experimental groups (wild-type, Tg2576 and Tg2576 + mAb) immunolabeled with anti-GFAP antibody (red) and Hoechst for nuclei visualization (blue); Scale bar 35 μm (c). Quantification of GFAP area covered by fluorescent signal/field of view (*p < 0.05 Tg2576 versus wild-type; *p < 0.05 Tg2576 versus Tg2576 + mAb; n = 8 slices/3 mice for each group; One-way ANOVA, post-hoc Bonferroni test) (d). e, f Representative images of retinal slices from animals of three experimental groups (wild-type, Tg2576 and Tg2576 + mAb) immunolabeled with anti-Iba1 antibody (green) and Hoechst for nuclei visualization (blue); Scale bar 35 μm (e). Quantification of number of microglia cells in the volume of each field of view (***p < 0.0005 wild-type versus Tg2576;***p < 0.0005 wild-type versus Tg2576 + mAb; n = 8 slices/3 mice for each group; One-way ANOVA post-hoc Bonferroni test). IL: inner layer; OL: outer layer