glun2a nmda receptor subunits  (Millipore)

Journal: International Journal of Molecular Sciences

Article Title: Use of Steaming Process to Improve Biochemical Activity of Polygonatum sibiricum Polysaccharides against D-Galactose-Induced Memory Impairment in Mice

doi: 10.3390/ijms231911220

Figure Lengend Snippet: Steamed PSPs ameliorate D-gal-induced hippocampal synaptic deficits. ( A – F ) The upper panels show representative blots of Arc/β-actin, GluA1/β-actin, GluA2/β-actin, GluN2A/β-actin, GluN2B/β-actin, p-GluN2B/GluN2B, and PSD95/β-actin, while the lower panel shows Arc/β-actin, GluA1/β-actin, GluA2/β-actin, GluN2A/β-actin, GluN2B/β-actin, p-GluN2B/GluN2B, and PSD95/β-actin quantification data ( n = 6). ( G ) Representative images of synaptic structure in hippocampal area CA1 (Magnification: ×5000). Red arrows indicate the synapses. ( H ) Number of synapses (12 μm 2 ) ( n = 5). Data are expressed as means ± SEM. * p < 0.05.

Article Snippet: Thereafter, protein samples were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis at 75 V for 15 min, followed by 115 V for 75 min before transferring at 300 mA onto a nitrocellulose membrane for 2 h. Membranes were blocked with 5% skim milk (dissolved in PBS containing 0.1% Tween-20) for 2 h at room temperature, washed three times with PBS for 10 min each, then incubated at 4 °C overnight in primary antibody against the α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor (AMPAR) subunits GluA1 (1:1000, CST) and GluA2 (1:1000, CST), the N-methyl-D-aspartate receptor (NMDAR) subunits GluN2A (1:1000, CST), GluN2B (1:1000, CST), and p-GluN2B (1:1000, CST), nuclear factor-erythroid factor 2-related factor 2 (Nrf2, 1:1000, ZEN BIO), heme oxygenase-1 (HO-1, 1:1000, ZEN BIO), NOD-like receptor protein 3 (NLRP3, 1:1000, CST), apoptosis-associated speck like protein (ASC, 1:1000, ZEN BIO), postsynaptic density protein 95 (PSD95, 1:1000, CST), activity-regulated cytoskeletal protein (Arc, 1:1000, Synaptic Systems), and β-actin (1:1000, CST).

Techniques: