Journal: Neuroscience Bulletin

Article Title: Rac1 Modulates Excitatory Synaptic Transmission in Mouse Retinal Ganglion Cells

doi: 10.1007/s12264-019-00353-0

Figure Lengend Snippet: Protein levels of mGluR1/5 and NMDAR subunits in retinal extracts from Rac1-cKO, Chat-cre+/–, and control mice. A Representative immunoblots showing the mGluR1 and mGluR5 protein levels. B Bar charts summarizing the average densitometry of immunoreactive bands of mGluR1 and mGluR5 expression. C Representative immunoblots showing the GluN1, GluN2A, and GluN2B protein levels. D Bar charts summarizing the average densitometry of immunoreactive bands of GluN1, GluN2A, and GluN2B expression. All the data are normalized to control. n = 6–7. *P < 0.05, ***P < 0.001 vs control.

Article Snippet: After blocking in 5% non-fat milk at room temperature for 2 h, the membranes were incubated overnight at 4°C with the following primary antibodies: polyclonal mouse anti-GluN1 (1:1000; BD Pharmingen, Franklin Lakes, NJ), polyclonal rabbit anti-GluN2A (1:200; Alomone Labs, Jerusalem, Israel), polyclonal rabbit anti-GluN2B (1:200; Alomone Labs), polyclonal rabbit anti-GluA1 (1:200; Alomone Labs), monoclonal rabbit anti-mGluR1 (1:1000; Cell Signaling Technology, Danvers, MA), monoclonal rabbit anti-mGluR5 (1:500; Abcam), polyclonal rabbit anti-glycine receptor alpha1+alpha2 (1:1000; Abcam), and polyclonal rabbit anti-GABA A receptor alpha1 (GABRA1) (1:1000; Abcam).

Techniques: Western Blot, Expressing

Journal: Frontiers in Neuroscience

Article Title: Additive Effects of Environmental Enrichment and Ketamine on Neuropathic Pain Relief by Reducing Glutamatergic Activation in Spinal Cord Injury in Rats

doi: 10.3389/fnins.2021.635187

Figure Lengend Snippet: Primary antibodies for western blotting.

Article Snippet: Rabbit anti-phospho-NMDAR2B Tyr 1472 (p-NR2B) , 1:1,000 , Merck Millipore, Darmstadt, Germany.

Techniques: Western Blot

Journal: Anesthesia & Analgesia

Article Title: Intrathecal Injection of the Peptide Myr-NR2B9c Attenuates Bone Cancer Pain Via Perturbing N-Methyl-D-Aspartate Receptor-PSD-95 Protein Interactions in Mice

doi: 10.1213/ane.0000000000000202

Figure Lengend Snippet: Figure 3. Myr-NR2B9c disrupted protein interactions between NR2B-containing NMDAR and PSD-95 in spinal cord. A, Western blot- ting showed immunoprecipitates (IP, 3 right lanes) of PSD-95 with NR2B from mice pretreated with vehicle (DMSO, 5 μL), Myr-NR2BAA (1 μg/5 μL) or Myr-NR2B9c peptide (1 μg/5 μL). The left lane showed normal lumbar spinal cord lysates acting as positive con- trol. B, Statistical analysis of the normalized OD (PSD-95/NR2B) among group vehicle, group Myr-NR2BAA, and group Myr-NR2B9c. Each group consisted of 3 mice. The data were expressed as the mean ± SD. *P < 0.01 versus mice treated with vehicle. IP = immu- noprecipitation; IB = immunoblotting.

Article Snippet: Filter membranes were blocked with 5% nonfat milk for 2 hours at room temperature and then incubated with the following primary antibodies, rabbit antiphospho-Tyr1472 NR2B (1:500, CST, Biotechnology), rabbit anti-PSD-95 (1:500; Biotechnology, Santa Cruz, CA), and rabbit anti-nNOS (1:1000; Abcam, United Kingdom) at 4°C overnight.

Techniques: Western Blot

Journal: Anesthesia & Analgesia

Article Title: Intrathecal Injection of the Peptide Myr-NR2B9c Attenuates Bone Cancer Pain Via Perturbing N-Methyl-D-Aspartate Receptor-PSD-95 Protein Interactions in Mice

doi: 10.1213/ane.0000000000000202

Figure Lengend Snippet: Figure 2. Changes in the spinal phospho-Tyr1472 NR2B, nNOS, and PSD-95 expression after tumor cells inoculation. Inoculation tumor cells into the intramedullary space of the right femur induced the upregulation of spinal phospho-Tyr1472 NR2B, nNOS, and PSD-95 expression from day 5 to day 14 postoperation. A, The representative bands of phospho-Tyr1472 NR2B, nNOS, and PSD-95 as assessed using western blotting at different time points after inoculation, respectively. β-actin was used as a loading control. Statistical analy- sis of the normalized OD (B) (p-NR2B/β-actin), (C) (nNOS/β-actin) and (D) (PSD-95/β-actin) between tumor-bearing and sham mice. Each group consisted of 3 mice. The data were expressed as the mean ± SD. *P < 0.01 versus sham mice.

Article Snippet: Filter membranes were blocked with 5% nonfat milk for 2 hours at room temperature and then incubated with the following primary antibodies, rabbit antiphospho-Tyr1472 NR2B (1:500, CST, Biotechnology), rabbit anti-PSD-95 (1:500; Biotechnology, Santa Cruz, CA), and rabbit anti-nNOS (1:1000; Abcam, United Kingdom) at 4°C overnight.

Techniques: Expressing, Western Blot, Control

Journal: Anesthesia & Analgesia

Article Title: Intrathecal Injection of the Peptide Myr-NR2B9c Attenuates Bone Cancer Pain Via Perturbing N-Methyl-D-Aspartate Receptor-PSD-95 Protein Interactions in Mice

doi: 10.1213/ane.0000000000000202

Figure Lengend Snippet: Figure 5. The effects of intrathecal administration of Myr-NR2B9c on the expression of phospho-Tyr1472 NR2B, nNOS, and PSD-95 in spinal cord. Tissue samples of L3-L5 spinal cord for western blotting were obtained from mice treated with Myr-NR2B9c before admin- istration and at 2, 4, 6, 8, 10, 12, and 24 hours after administra- tion. The expression levels before administration were regarded as baseline values. A, The representative bands of phospho-Tyr1472 NR2B, nNOS, and PSD-95 detected by western blotting at different time points after administration of NR2B9c, respectively. β-actin was used as a loading control. Statistical analysis of the normalized OD (B) (p-NR2B/β-actin), (C) (nNOS/β-actin) and (D) (PSD-95/β-actin). Intrathecal injection of Myr-NR2B9c decreased the expression of phospho-Tyr1472 NR2B, nNOS, and PSD-95 in different manners. Each group consisted of 3 mice. The data were expressed as the mean ± SD. *P < 0.01 versus baseline.

Article Snippet: Filter membranes were blocked with 5% nonfat milk for 2 hours at room temperature and then incubated with the following primary antibodies, rabbit antiphospho-Tyr1472 NR2B (1:500, CST, Biotechnology), rabbit anti-PSD-95 (1:500; Biotechnology, Santa Cruz, CA), and rabbit anti-nNOS (1:1000; Abcam, United Kingdom) at 4°C overnight.

Techniques: Expressing, Western Blot, Control, Injection