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Fig. 4 | Acta Neuropathologica Communications

Fig. 4

From: Systemic delivery of a specific antibody targeting the pathological N-terminal truncated tau peptide reduces retinal degeneration in a mouse model of Alzheimer’s Disease

Fig. 4

Immunotherapy with 12A12mAb mitigates the AD-associated synaptic and apoptotic changes and prevents the microtubule destabilization in retinas from diseased animals. Western blotting analyses (a) were carried out on equal amounts of total protein extract (50 µg) from retinas of animals of three experimental groups (wild-type, Tg2576 and Tg2576 + mAb). Immunoblots were probed with antibodies against several pre- and postsynaptic proteins—including the N-Methyl-D-aspartate (NMDA) receptor subunits 1 (NR1), synapsin I, synaptophysin, syntaxin 1, SNAP25, α-synuclein and the active (cleaved) form of caspase-6 (Asp162). Data were quantified for molecular weight size ranges for each antibody and normalized to β-actin which was used as loading control. Relative intensity of each protein was calculated and semi-quantitative densitometric analysis (n = 7) is shown (b). Arrows on the right side indicate the molecular weight (kDa) of bands calculated from migration of standard proteins. Statistically significant differences (see details in the main text) 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). The functional integrity of axonal track was evaluated by probing the immunoblots with antibodies against the acetyl- and tyrosinylated-α-tubulin, as markers for stable and unstable/dynamic microtubule respectively (c). Data were quantified for molecular weight size ranges for each antibody and normalized to β-actin which was used as loading control. Relative intensity of each protein was calculated and semi-quantitative densitometric analysis (n = 6) is shown (d). Arrows on the right side indicate the molecular weight (kDa) of bands calculated from migration of standard proteins. Statistically significant differences (see details in the main text) 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)

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