p nr2b  (Proteintech)

Journal: Addiction Biology

Article Title: NMDA receptor within nucleus accumbens shell regulates propofol self‐administration through D1R/ERK/CREB signalling pathway

doi: 10.1111/adb.13401

Figure Lengend Snippet: Images and plots of western blotting analysis of D1R/ERK/CREB signalling pathway in the NAc after the pretreatment of D‐AP5 in bilateral NAsh in propofol self‐administrated rats. The expressions of D1R (A, p < 0.001, n = 7, ANOVA), p‐ERK1/2 (B, p = 0.001, n = 7, Kruskal–Wallis) and p‐CREB (D, p = 0.001, n = 7, ANOVA) in the NAc were statistically significantly increased in a dose‐dependent manner by D‐AP5 pretreatment. Compared with the vehicle group, the pairwise analysis showed the difference reached a statistical significance at the dose of 2.0–4.0 μg/0.3 μL/site for D1Rs (2.0 group p < 0.001, 4.0 group p < 0.001, Dunnett) and p‐CREB (p‐CREB, 2.0 group p = 0.003, 4.0 group p = 0.001, Dunnett), and at the dose of 4.0 μg/0.3 μL/site for p‐ERK1/2 ( p = 0.001, Dunn). D‐AP5 pretreatment failed to affect the expression of t‐ERK1/2 (C, p = 0.996, n = 6, ANOVA) and t‐CREB (E, p = 0.798, n = 6, ANOVA) in the NAc. p‐ERK1/2 = phosphorylated ERK1/2, t‐ERK1/2 = total ERK1/2, p‐CREB = phosphorylated CREB, t‐NR2B = total CREB. The normally distributed data were analysed with one‐way ANOVA, and Dunnett's post hoc test was conducted for multiple comparisons; otherwise, the data that did not meet the criteria of normal distribution were analysed by Kruskal–Wallis test with Dunn's post hoc analysis for multiple comparisons. Compared with the vehicle group, ** p < 0.01, *** p < 0.001.

Article Snippet: The protein was transferred to polyvinylidene fluoride (PVDF) membranes, and the non‐specific binding sites were blocked with 5% skim milk at room temperature for 2 h. After that, the bands were incubated in primary antibody [NR2A, p‐NR2A (Tyr1246), NR2B, p‐NR2B (Tyr1472), p‐ERK and p‐CREB, rabbit, 1:1000, Cell Signaling Technology, Danvers, MA, United States; D1R, ERK and CREB, mouse, 1:1000, Santa Cruz Biotechnology, Texas, United States] at 4°C overnight and in the secondary antibody (goat anti‐rabbit or goat anti‐mouse, 1:3000, Proteintech, Chicago, United States) that diluted with tris‐buffered saline (TBST) at room temperature for 2 h. We adopted glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) (mouse, 1:10000, Proteintech, Chicago, United States) as the internal control.

Techniques: Western Blot, Expressing

Journal: The Journal of International Medical Research

Article Title: Hydrogen treatment reduces electroencephalographic activity and neuronal death in rats with refractory status epilepticus by inhibiting membrane NR2B phosphorylation and oxidative stress

doi: 10.1177/03000605241235589

Figure Lengend Snippet: Effects of hydrogen-rich saline (HRS) treatment on hippocampal cell membrane N-methyl-D-aspartate receptor subtype 2B (NR2B) expression. (a) Membrane NR2B expression was measured by western blot at different time points after refractory status epilepticus (RSE) induction ( n = 3). (c) Membrane NR2B expression at different time points after RSE induction in the RSE + saline and RSE + HRS groups and (b, d) relative protein levels in each group ( n = 3). Data are displayed as the mean ± standard error, * P < 0.05.

Article Snippet: After being blocked for 1 hour, the membranes were incubated overnight at 4°C with the appropriate primary antibodies—NR2B (ab28373, 1:500; Abcam, Cambridge, UK), phosphorylated (p)-NR2B (ab81271, 1:300; Abcam), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; CW0100 M, 1:2000; CoWin Biosciences, Taizhou, China)—followed by a 2-hour incubation at room temperature with secondary antibodies (goat anti-rabbit IgG, CW0103S, and goat anti-mouse IgG, CW0102S; both CoWin Biosciences).

Techniques: Saline, Membrane, Expressing, Western Blot

Journal: The Journal of International Medical Research

Article Title: Hydrogen treatment reduces electroencephalographic activity and neuronal death in rats with refractory status epilepticus by inhibiting membrane NR2B phosphorylation and oxidative stress

doi: 10.1177/03000605241235589

Figure Lengend Snippet: Effects of hydrogen-rich saline (HRS) treatment on hippocampal cell membrane N-methyl-D-aspartate receptor subtype 2B (NR2B) phosphorylation. (a) NR2B phosphorylation was measured by western blot at different time points after refractory status epilepticus (RSE) induction ( n = 3). (c) NR2B phosphorylation in the RSE + saline and RSE + HRS groups at different time points after RSE induction and (b, d) relative protein levels in each group ( n = 3). Data are displayed as the mean ± standard error, * P < 0.05.

Article Snippet: After being blocked for 1 hour, the membranes were incubated overnight at 4°C with the appropriate primary antibodies—NR2B (ab28373, 1:500; Abcam, Cambridge, UK), phosphorylated (p)-NR2B (ab81271, 1:300; Abcam), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; CW0100 M, 1:2000; CoWin Biosciences, Taizhou, China)—followed by a 2-hour incubation at room temperature with secondary antibodies (goat anti-rabbit IgG, CW0103S, and goat anti-mouse IgG, CW0102S; both CoWin Biosciences).

Techniques: Saline, Membrane, Western Blot