primary antibody [nr2a, p‐nr2a (tyr1246), nr2b, p‐nr2b (tyr1472), p‐erk and p‐creb  (Cell Signaling Technology Inc)

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: Translational Neuroscience

Article Title: Vitamin D3 reduces hippocampal NR2A and anxiety in nicotine withdrawal mice

doi: 10.1515/tnsci-2020-0166

Figure Lengend Snippet: Primers used for RT-PCR analysis

Article Snippet: NR2A primary antibody (19953-1-AP, 1:800) and GAPDH primary antibody (10494-1-AP, 1:10,000) were purchased from proteintech.

Techniques: Sequencing

Journal: Translational Neuroscience

Article Title: Vitamin D3 reduces hippocampal NR2A and anxiety in nicotine withdrawal mice

doi: 10.1515/tnsci-2020-0166

Figure Lengend Snippet: The hippocampal upregulation of NR2A level induced by nicotine withdrawal is corrected in mice fed with the vitamin D3-supplemented diet. NR2A mRNA level (a) and protein level (c) were significantly decreased after receiving vitamin D3 supplement, compared to nicotine-treated mice ( n = 8). There was a slight but significant decrease in hippocampal α7 nAChR in the nicotine-treated and vitamin D3-supplemented mice, compared with the vehicle group ( n = 8), both in mRNA expression (b) and in protein level (d). GAPDH was used as a reference protein ( n = 8). Data were expressed as mean ± SEM. Compared with the vehicle group * P < 0.05, ** P < 0.01; compared with the Nic group, # P < 0.05, ## P < 0.01.

Article Snippet: NR2A primary antibody (19953-1-AP, 1:800) and GAPDH primary antibody (10494-1-AP, 1:10,000) were purchased from proteintech.

Techniques: Expressing

Journal: Experimental neurology

Article Title: Neurosteroid allopregnanolone reduces ipsilateral visual cortex potentiation following unilateral optic nerve injury

doi: 10.1016/j.expneurol.2018.05.005

Figure Lengend Snippet: VGlut2 expression in the left and right visual cortex

Article Snippet: Membranes were blocked in 5% non-fat milk probed with primary antibodies against VGlut2 (1:5000, AB2251, Millipore), GABA A receptor δ-subunit (1:500, AB9752, Millipore), NR2A (1:1000, AB1555P, Millipore).

Techniques: Expressing