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. Author manuscript; available in PMC: 2019 Oct 1.
Published in final edited form as: Acta Neuropathol. 2018 Jul 11;136(4):589–605. doi: 10.1007/s00401-018-1886-z

Fig. 3. Bidirectional redistribution of AβOs caused by αSyn in APP mice.

Fig. 3

(a) Detection of oligomeric amyloid conformers in extracellular-enriched (EC) and membrane-enriched (MB) lysates from APP/αSyn, APP and APP/αSyn-KO mice by dot blot analysis using A11 and OC antibodies. 6E10 was also used to measure APP/Aβ abundance and actin was used as internal control. (n = 6 animals/age/genotype). Note that only 3 (out of 6) APP/αSyn specimens were adsorbed onto the presented nitrocellulose membranes shown for A11, OC and Actin. Grey and teal rectangles correspond to WT and αSyn control lysates respectively. Note that Actin is not present in extracellular-enriched lysates as expected. (b) Normalized abundance of oligomeric species indicated a bidirectional redistribution of A11 and OC conformers in EC and MB extracts of APP animals. (Histograms represent the mean ± S.D.; One-way ANOVA [F(2,18)A11-EC = 12.4110, P = 0.0008, F(2,18)OC-EC = 22.6278, P < 0.0004, F(2,18)6E10-EC = 8.5928, P = 0.0037 and F(2,18)A11-MB = 5.8887, P = 0.0139, F(2,18)OC-MB = 53.8985, P < 0.0001, F(2,18)6E10-MB = 5.131, P = 0.0123 respectively] followed by Student t test with Bonferroni correction; ★P < 0.05 vs. 6-month-old APP mice, n = 6 animals/age/genotype). (c) Representative confocal images of hippocampi labeled for αSyn (magenta; 4D6 antibody) and prefibrillar AβOs (green, OC antibody) from 6-month-old APP/αSyn, APP and APP/αSyn-KO mice (n = 6 animals/age/genotype). Arrows indicate OC-positive Aβ deposits in APP and APP/αSyn-KO mice. Note the synaptic pattern of OC+ AβOs in APP/αSyn mice (empty white box). Scale bars = 200 μm. (d) Detection of oligomeric αSyn in intracellular enriched (IC) lysates by dot blot analysis using Syn33 and F8H7 antibodies. 4D6 and LB509 were used to detect total or human αSyn respectively and actin was used as an internal control. (n = 8 animals/age/genotype). Grey and teal rectangles correspond to WT and αSyn control lysates respectively. (e) Quantitation Syn33 oligomeric species in IC extracts. Histograms represent the mean ± S.D.; One-way ANOVA [F(4,40) = 304.3331, P < 0.0001 for Syn33] followed by Student t test with Bonferroni correction; ★P < 0.05 vs. 6-month-old APP mice, ☆P < 0.0001 vs. αSyn. (n = 8 animals/age/genotype)