Journal: Neurotherapeutics
Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model
doi: 10.1016/j.neurot.2025.e00739
Figure Lengend Snippet: The ErbB4 small molecule agonist modulates DOCK3 to ameliorate hippocampal neuronal injury under AD pathological conditions in vitro. (A–H) After a 12 h exposure to 10 μM Aβ, the cells were transfected with either 50 nM si-NC or si-DOCK3 for 24 h. This was followed by a combined treatment with C11H7BrO3 at a concentration of 10 nM for an additional 24 h. (A) Protein levels of the ErbB4 pathway-related proteins in hippocampal neurons were assessed using Western blot analysis. (B) Statistical representation of these protein levels was generated in a chart. (C) Protein levels of DOCK3, CREB, SIRT3 in hippocampal neurons were assessed using Western blot analysis. (D) Statistical representation of these protein levels was generated in a chart. (E) Protein levels of the synapse proteins SYT1, GAP43, SYP, and PSD95 in hippocampal neurons were assessed using Western blot analysis. (F) Statistical representation of these protein levels was generated in a chart. (G) Levels of oxidative stress indicators in hippocampal neurons, including T-SOD, GSH, GSH-Px, and MDA. (H) Intracellular ATP and extracellular ATP (eATP) levels in hippocampal neurons. (I–J). Representative images and quantitative analysis of mitochondrial signaling and membrane potential in hippocampal neurons. (K) The mitochondrial homeostasis function of hippocampal neurons involved the measurement of Mito-Tracker Green and TMRE levels. Tukey multiple comparative analyses were conducted, with data reported as means ± SEM. Significance levels were denoted as ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Scale bar = 20 μm. A minimum sample size of n = 5–6 per group was utilized.
Article Snippet: Specifically, the fixed cells were treated with mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz biotech, USA) in combination with the following antibodies: rabbit anti-SYT1 (1:200, GB111130-100, Servicebio, Wuhan, China), rabbit anti-GAP43 antibody (1:1000, GB15094-100, Servicebio, Wuhan, China), rabbit anti- p -ErbB4 (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP (1:200, AF8091, Bioss, Beijing, China), rabbit anti-PSD95 (1:200, GB11277-100, Servicebio, Wuhan, China) and rabbit anti-DOCK3 (1:200, 20683-1-AP, proteintech, Wuhan, China).
Techniques: In Vitro, Transfection, Concentration Assay, Western Blot, Generated, Membrane