Journal: Alzheimer's & dementia : the journal of the Alzheimer's Association
Article Title: Reduced synaptic proteins and SNARE complexes in Down syndrome with Alzheimer's disease and the Dp16 mouse Down syndrome model: Impact of APP gene dose.
doi: 10.1002/alz.12835
Figure Lengend Snippet: FIGURE 2 Reduction of SNARE complexes in AD-DS, AD, and Dp16 samples. Syntaxin 1A was present in a 35 kD monomeric form and in a 73 kD trimeric complex with SNAP25 and synaptobrevin 2 as well as other high molecular weight forms. A, D, G, Unboiled AD-DS (A), DS (D), and AD (G) SDS samples were loaded on SDS-PAGE and probed with the syntaxin 1A antibody. A boiled sample was also loaded as a control to demonstrate disruption of the complexes with heating. The arrow points to the 73 kD trimeric band. B, E, H, Profiles of relative intensities of the ladder of complexes from the representative samples marked with asterisks; the heaviest complexes are represented at the left end of the x-axis. C, F, I, Quantitation and statistical analysis of the levels of 73 kD trimeric SNARE complex in AD-DS and DS samples. Mann–Whitney test; *P < 0.05, **P < 0.01. J, Unboiled 2N, Dp16, and Dp16: App++−samples from mice aged 9 to 10 months were loaded on SDS-PAGE and probed with the syntaxin 1A antibody. Arrows point to the major 73 kD, 100–130 kD, and > 180 kD bands. K–M, Quantitation and statistical analysis of the levels of 73 kD, 100–130 kD and > 180 kD bands. One-way analysis of variance test followed by Newman–Keuls multiple comparison test. n = 3 mice for each category; *P < 0.05. C/, control for; AD-DS, Alzheimer’s disease with Down syndrome; DS, Down syndrome without Alzheimer’s disease; AD, Alzheimer’s disease; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis
Article Snippet: 3.1 Methods 3.1.1 Antibodies Mouse antibody anti-syntaxin 1A (1:5000; S0664; HPC-1) and rabbit antibody anti-APP (1:3000; A8717) were obtained from MilliporeSigma; rabbit antibody anti-SNAP25 (1:2000; 14903-1-AP), antiNPTX2 (10889-1-AP), mouse antibodies anti-synaptophysin (1:5000; 67864-1-Ig), anti-β-actin (1:10,000; 60008-1-Ig) were from Proteintech.
Techniques: High Molecular Weight, SDS Page, Control, Disruption, Quantitation Assay, MANN-WHITNEY, Comparison, Polyacrylamide Gel Electrophoresis