elisa  (Cell Signaling Technology Inc)

Journal: iScience

Article Title: Peripheral amylin modulation rebalances brain glycolysis and Tau-Ser214 phosphorylation via cAMP-PKA signaling

doi: 10.1016/j.isci.2026.115157

Figure Lengend Snippet: Effects of circulating amylin modulation on AD-like pathology in vivo and in human amylin-treated neonatal neurons (A–D) Heat maps comparing the brain tissue levels of pT231-tau, tau, Aβ 40 , and Aβ 42 in the same mice as in , , , , and (i.e., hA ON vs. hA i−OFF mice and hA OFF vs. hA i−ON mice). (E) Aβ immunofluorescence signal for Aβ (green) of cultured primary neurons incubated with human amylin (hAmylin; 1 μM for 4 h) or under control conditions (control). MAP2 (blue) was used to identify the neurons. n = 86 neurons from 3 primary neonatal rat neuron cultures. (F) Total Aβ level secreted in the culture media by primary neurons incubated with human amylin (hAmylin; 1 μM for 2 h) or under control conditions (control). Aβ was enriched by immunoprecipitation and measured by Western blot. (G) pTau and total Tau lysates from primary neurons incubated with human amylin (hAmylin; 1 μM for 2 h) or under control conditions (control). pTau and Tau were enriched by immunoprecipitation and measured by western blot. Individual data points and mean ± s.e.m for three neuronal cultures. (H) Effect of amylin on pS214-tau in cultured murine astrocytes. Cells were incubated for 30 min under control conditions or in the presence of human amylin (15 μM) with and without the amylin receptor antagonist AC187 (10 μM) or PKA inhibitor H-89 (10 μM). pS214-tau was measured by ELISA. Individual data points and mean ± s.e.m for three cell cultures. (I) Representative images of IHC analyses of cortical slices from hA ON mice stained for amylin (top) and for total Aβ (bottom) ( n = 5 slices/mouse from n = 3 mice). Statistical analyses were performed using one-way ANOVA followed by Dunnett’s multiple-comparisons test (H) and two-tail t test (E–G). See also .

Article Snippet: pS214-tau , Cell Signaling , 44177.

Techniques: In Vivo, Immunofluorescence, Cell Culture, Incubation, Control, Immunoprecipitation, Western Blot, Enzyme-linked Immunosorbent Assay, Staining

Journal: iScience

Article Title: Peripheral amylin modulation rebalances brain glycolysis and Tau-Ser214 phosphorylation via cAMP-PKA signaling

doi: 10.1016/j.isci.2026.115157

Figure Lengend Snippet: Pancreatic amylin regulation reduces amylin-cAMP-PKA overactivation and tau-Aβ coupling: molecular evidence (A and B) Representative confocal microscopy images comparing neuronal pS214-tau in cortical sections from hA ON vs. hA OFF mice ( n = 5 slices/mouse from n = 3 mice/group). (C–F) In the same mice as in , , , , , , and , average brain amylin level vs. average pS214-tau (C) and average brain tissue PKA activity vs. average pS214-tau (D) along with correlation analyses between brain tissue Aβ 42 and pT31-tau in hA ON vs. hA i−OFF mice (E) and hA OFF vs. hA i−ON mice (F). (G) Heat maps comparing the brain tissue levels of inflammatory cytokines (IL-1β, IL-6, TNF-α, IFN-γ, and CXCL1) in hA i−OFF mice vs. hA ON littermates. (H) Same as in (G) for hA i−ON mice compared to hA OFF littermates. (I) In same mice, principal component (PC) analysis of amylin, AD markers (Aβ 40, Aβ 42 , pTau, and total tau), neuroinflammation (IL-1β, IL-6, TNF-α, IFN-γ, and CXCL1), and metabolic covariates (glucose, G6P, cAMP, and ATF4). (J) PC loading plot suggesting that clustering is largely driven by contributions of amylin, cAMP and Aβ covariates to the main principal component (PC1), whereas G6P and tau pathology drive the variation on PC2. (K) Schematic diagram (using BioRender): toggling amylin secretion under pre-diabetic stress bidirectionally regulates brain glucose metabolism and memory. Excess amylin amplifies amylin-cAMP-PKA signaling, suppresses astrocytic glycolysis and promotes tau phosphorylation, tau-Aβ coupling and neuroinflammation, whereas restricting brain amylin preserves metabolic function and memory.

Article Snippet: pS214-tau , Cell Signaling , 44177.

Techniques: Confocal Microscopy, Activity Assay, Phospho-proteomics