anti p nr2b  (Cell Signaling Technology Inc)

Journal: Brain research bulletin

Article Title: The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning.

doi: 10.1016/j.brainresbull.2025.111400

Figure Lengend Snippet: Fig. 1. Effects of hypoxia preconditioning on GluN2B, pY1252 GluN2B, and pY1336 GluN2B levels in the mouse hippocampus. A: The mRNA expression level of GluN2B (n = 6 per group);B: Representative Western blotting results of GluN2B, pY1252 GluN2B, and pY1336 GluN2B (n = 6 per group);C-E: Semi-quantitative analysis showing the protein changes in GluN2B, pY1252 GluN2B, and pY1336 GluN2B;F-H: Immunofluorescence imaging and relative fluorescence intensity of GluN2B, pY1252 GluN2B, and pY1336 GluN2B in the mouse hippocampus on day 0 (n = 5 per group). Immunofluorescence imaging (300 dpi) was acquired using a laser scanning confocal microscope (A1; Nikon) with 40x magnification. Scale bar= 100μM.* p﹤0.05 versus N group; # p﹤0.05 versus H group. N: Normoxia; H: Hypoxia; HPC: Hypoxia Preconditioning.

Article Snippet: The membranes were incubated overnight with the following primary antibodies: (GluN2B (Cell Signaling Technology, #4207), p-Y1336 GluN2B (Phosphosolutions, #p1516–1336), p-Y1252 GluN2B (Phosphosolutions, #p1516–1252), spectrin (Santa Cruz Biotechnology, sc-48382), caspase-3(Cell Signaling Technology, #9662), β-actin (Santa Cruz Biotechnology, sc-47778), PSD95 antibody (Cell Signaling Technology, #3450),GluN1 antibody (Cell Signaling Technology, #5704),GluN2A antibody (Cell Signaling Technology, #4205),EEA1 antibody (Cell Signaling Technology, #2411),Rab11 antibody (Cell Signaling Technology, #5589), p97 ATPase antibody (Cell Signaling Technology, #2649)), and then incubated with secondary antibodies for 1 h. The blots were detected using ECL ultrasensitive luminescence solution (Zhang et al., 2019).

Techniques: Expressing, Western Blot, Immunofluorescence, Imaging, Fluorescence, Microscopy

Journal: Brain research bulletin

Article Title: The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning.

doi: 10.1016/j.brainresbull.2025.111400

Figure Lengend Snippet: Fig. 3. Effects of hypoxia preconditioning on the distribution of GluN2B, pY1252 GluN2B, and pY1336 GluN2B in synaptic and extrasynaptic components of the mouse hippocampus. A: Representative Western blotting results showing GluN2B, pY1252 GluN2B, and pY1336 GluN2B in the synaptic member (TxP); B-D: Semi- quantitative analysis showing the protein changes in GluN2B, pY1252 GluN2B, and pY1336 GluN2B in the TxP (n = 6 per group); E: Representative Western blotting results showing GluN2B, pY1252 GluN2B, and pY1336 GluN2B in the extra synaptic membrane (TxS); F-H: Semi-quantitative analysis showing the protein changes in GluN2B, pY1252 GluN2B, and pY1336 GluN2B in TxP (n = 6 per group); * p﹤0.05 versus N group; # p﹤0.05 versus H group. N: Normoxia; H: Hypoxia; HPC: Hypoxia Preconditioning.

Article Snippet: The membranes were incubated overnight with the following primary antibodies: (GluN2B (Cell Signaling Technology, #4207), p-Y1336 GluN2B (Phosphosolutions, #p1516–1336), p-Y1252 GluN2B (Phosphosolutions, #p1516–1252), spectrin (Santa Cruz Biotechnology, sc-48382), caspase-3(Cell Signaling Technology, #9662), β-actin (Santa Cruz Biotechnology, sc-47778), PSD95 antibody (Cell Signaling Technology, #3450),GluN1 antibody (Cell Signaling Technology, #5704),GluN2A antibody (Cell Signaling Technology, #4205),EEA1 antibody (Cell Signaling Technology, #2411),Rab11 antibody (Cell Signaling Technology, #5589), p97 ATPase antibody (Cell Signaling Technology, #2649)), and then incubated with secondary antibodies for 1 h. The blots were detected using ECL ultrasensitive luminescence solution (Zhang et al., 2019).

Techniques: Western Blot, Membrane

Journal: Brain research bulletin

Article Title: The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning.

doi: 10.1016/j.brainresbull.2025.111400

Figure Lengend Snippet: Fig. 5. Effects of hypoxia preconditioning on the protein levels of GluN2B, pY1252 GluN2B, and pY1336 GluN2B in HT22 cells. A: Representative Western blotting results showing GluN2B, pY1252 GluN2B, and pY1336 GluN2B; B-D: Semi-quantitative analysis showing the protein changes in GluN2B, pY1252 GluN2B, and pY1336 GluN2B (n = 3 per group); E-G: Immunofluorescence imaging and relative fluorescence intensities of GluN2B, pY1252 GluN2B, and pY1336 GluN2B in HT22 cells(n = 5 per group). Immunofluorescence imaging (300 dpi) was acquired using a laser scanning confocal microscope (A1; Nikon) with 200x magnification. Scale bar= 50μM. * p﹤0.05 versus N group; # p﹤0.05 versus H group. N: Normoxia; H: Hypoxia; HPC: Hypoxia Preconditioning.

Article Snippet: The membranes were incubated overnight with the following primary antibodies: (GluN2B (Cell Signaling Technology, #4207), p-Y1336 GluN2B (Phosphosolutions, #p1516–1336), p-Y1252 GluN2B (Phosphosolutions, #p1516–1252), spectrin (Santa Cruz Biotechnology, sc-48382), caspase-3(Cell Signaling Technology, #9662), β-actin (Santa Cruz Biotechnology, sc-47778), PSD95 antibody (Cell Signaling Technology, #3450),GluN1 antibody (Cell Signaling Technology, #5704),GluN2A antibody (Cell Signaling Technology, #4205),EEA1 antibody (Cell Signaling Technology, #2411),Rab11 antibody (Cell Signaling Technology, #5589), p97 ATPase antibody (Cell Signaling Technology, #2649)), and then incubated with secondary antibodies for 1 h. The blots were detected using ECL ultrasensitive luminescence solution (Zhang et al., 2019).

Techniques: Western Blot, Immunofluorescence, Imaging, Fluorescence, Microscopy

Journal: Brain research bulletin

Article Title: The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning.

doi: 10.1016/j.brainresbull.2025.111400

Figure Lengend Snippet: Fig. 1. Effects of hypoxia preconditioning on GluN2B, pY1252 GluN2B, and pY1336 GluN2B levels in the mouse hippocampus. A: The mRNA expression level of GluN2B (n = 6 per group);B: Representative Western blotting results of GluN2B, pY1252 GluN2B, and pY1336 GluN2B (n = 6 per group);C-E: Semi-quantitative analysis showing the protein changes in GluN2B, pY1252 GluN2B, and pY1336 GluN2B;F-H: Immunofluorescence imaging and relative fluorescence intensity of GluN2B, pY1252 GluN2B, and pY1336 GluN2B in the mouse hippocampus on day 0 (n = 5 per group). Immunofluorescence imaging (300 dpi) was acquired using a laser scanning confocal microscope (A1; Nikon) with 40x magnification. Scale bar= 100μM.* p﹤0.05 versus N group; # p﹤0.05 versus H group. N: Normoxia; H: Hypoxia; HPC: Hypoxia Preconditioning.

Article Snippet: The membranes were incubated overnight with the following primary antibodies: (GluN2B (Cell Signaling Technology, #4207), p-Y1336 GluN2B (Phosphosolutions, #p1516–1336), p-Y1252 GluN2B (Phosphosolutions, #p1516–1252), spectrin (Santa Cruz Biotechnology, sc-48382), caspase-3(Cell Signaling Technology, #9662), β-actin (Santa Cruz Biotechnology, sc-47778), PSD95 antibody (Cell Signaling Technology, #3450),GluN1 antibody (Cell Signaling Technology, #5704),GluN2A antibody (Cell Signaling Technology, #4205),EEA1 antibody (Cell Signaling Technology, #2411),Rab11 antibody (Cell Signaling Technology, #5589), p97 ATPase antibody (Cell Signaling Technology, #2649)), and then incubated with secondary antibodies for 1 h. The blots were detected using ECL ultrasensitive luminescence solution (Zhang et al., 2019).

Techniques: Expressing, Western Blot, Immunofluorescence, Imaging, Fluorescence, Microscopy

Journal: Brain research bulletin

Article Title: The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning.

doi: 10.1016/j.brainresbull.2025.111400

Figure Lengend Snippet: Fig. 3. Effects of hypoxia preconditioning on the distribution of GluN2B, pY1252 GluN2B, and pY1336 GluN2B in synaptic and extrasynaptic components of the mouse hippocampus. A: Representative Western blotting results showing GluN2B, pY1252 GluN2B, and pY1336 GluN2B in the synaptic member (TxP); B-D: Semi- quantitative analysis showing the protein changes in GluN2B, pY1252 GluN2B, and pY1336 GluN2B in the TxP (n = 6 per group); E: Representative Western blotting results showing GluN2B, pY1252 GluN2B, and pY1336 GluN2B in the extra synaptic membrane (TxS); F-H: Semi-quantitative analysis showing the protein changes in GluN2B, pY1252 GluN2B, and pY1336 GluN2B in TxP (n = 6 per group); * p﹤0.05 versus N group; # p﹤0.05 versus H group. N: Normoxia; H: Hypoxia; HPC: Hypoxia Preconditioning.

Article Snippet: The membranes were incubated overnight with the following primary antibodies: (GluN2B (Cell Signaling Technology, #4207), p-Y1336 GluN2B (Phosphosolutions, #p1516–1336), p-Y1252 GluN2B (Phosphosolutions, #p1516–1252), spectrin (Santa Cruz Biotechnology, sc-48382), caspase-3(Cell Signaling Technology, #9662), β-actin (Santa Cruz Biotechnology, sc-47778), PSD95 antibody (Cell Signaling Technology, #3450),GluN1 antibody (Cell Signaling Technology, #5704),GluN2A antibody (Cell Signaling Technology, #4205),EEA1 antibody (Cell Signaling Technology, #2411),Rab11 antibody (Cell Signaling Technology, #5589), p97 ATPase antibody (Cell Signaling Technology, #2649)), and then incubated with secondary antibodies for 1 h. The blots were detected using ECL ultrasensitive luminescence solution (Zhang et al., 2019).

Techniques: Western Blot, Membrane

Journal: Brain research bulletin

Article Title: The decrease of GluN2B and its phosphorylation at Tyr-1336 in extrasynaptic subunits is associated with neuroprotection induced by hypoxia preconditioning.

doi: 10.1016/j.brainresbull.2025.111400

Figure Lengend Snippet: Fig. 5. Effects of hypoxia preconditioning on the protein levels of GluN2B, pY1252 GluN2B, and pY1336 GluN2B in HT22 cells. A: Representative Western blotting results showing GluN2B, pY1252 GluN2B, and pY1336 GluN2B; B-D: Semi-quantitative analysis showing the protein changes in GluN2B, pY1252 GluN2B, and pY1336 GluN2B (n = 3 per group); E-G: Immunofluorescence imaging and relative fluorescence intensities of GluN2B, pY1252 GluN2B, and pY1336 GluN2B in HT22 cells(n = 5 per group). Immunofluorescence imaging (300 dpi) was acquired using a laser scanning confocal microscope (A1; Nikon) with 200x magnification. Scale bar= 50μM. * p﹤0.05 versus N group; # p﹤0.05 versus H group. N: Normoxia; H: Hypoxia; HPC: Hypoxia Preconditioning.

Article Snippet: The membranes were incubated overnight with the following primary antibodies: (GluN2B (Cell Signaling Technology, #4207), p-Y1336 GluN2B (Phosphosolutions, #p1516–1336), p-Y1252 GluN2B (Phosphosolutions, #p1516–1252), spectrin (Santa Cruz Biotechnology, sc-48382), caspase-3(Cell Signaling Technology, #9662), β-actin (Santa Cruz Biotechnology, sc-47778), PSD95 antibody (Cell Signaling Technology, #3450),GluN1 antibody (Cell Signaling Technology, #5704),GluN2A antibody (Cell Signaling Technology, #4205),EEA1 antibody (Cell Signaling Technology, #2411),Rab11 antibody (Cell Signaling Technology, #5589), p97 ATPase antibody (Cell Signaling Technology, #2649)), and then incubated with secondary antibodies for 1 h. The blots were detected using ECL ultrasensitive luminescence solution (Zhang et al., 2019).

Techniques: Western Blot, Immunofluorescence, Imaging, Fluorescence, Microscopy

Journal: The Journal of Headache and Pain

Article Title: Alleviation of migraine related pain and anxiety by inhibiting calcium-stimulating AC1-dependent CGRP in the insula of adult rats

doi: 10.1186/s10194-024-01778-3

Figure Lengend Snippet: Enhanced NMDA receptors-mediated currents and their expression in the IC. A The synaptic input-output curve in slices from IS rats was steeper than in those from PBS group (IS: n =10 neurons/6 rats, PBS: n =11 neurons/5 rats; two-way ANOVA with Bonferroni post hoc). B I-V curves of NMDAR-EPSCs of IC pyramidal neurons recorded at holding potentials ranging from − 80 to + 40 mV in PBS and IS rats (IS: n =12 neurons/5 rats, PBS: n =10 neurons/5 rats; two-way ANOVA with Bonferroni post hoc). C A timeline plot of one representative neuron in IC showing the contribution of NMDA GluN2A and GluN2B -mediated currents (left). PEAQX (0.4 μM): selective GluN2A antagonist, Ro25-6981 (3 μM): selective GluN2B antagonist. Summarized data showing the contribution of GluN2A (two-tailed independent sample t-test) and GluN2B (two-tailed independent sample rank sum test) -mediated currents in IC neurons (IS: n =9 neurons/6 rats, PBS: n =6 neurons/5 rats; right). D Representative western blotting for GluN2B and p-GluN2B-S1303 in the IC from IS and PBS rats. E The total protein levels of GluN2B, p-GluN2B-S1303 significantly enhanced in IS rats ( n =5 rats/group; two-tailed independent sample t-test). F The phosphorylation level of GluN2B, p-GluN2B-S1303, significantly enhanced in IS rats ( n =5 rats/group; two-tailed independent sample t-test). All data are presented as the mean ± SEM (# P <0.05, ## P <0.01, IS vs. PBS)

Article Snippet: The PVDF membranes were blocked with a quick blocking solution (Yang Guang Bio, China) for one minute, and then incubated with the following antibodies: CGRP (1:1000, cell signaling technology, #14959, USA), GluN2B (1:2000, Proteintech, 21920-1-AP, China), p-GluN2B-S1303 (1:2000, abcam, ab81271, UK), AC1 (1:1000, abcam, ab69597, UK), β-actin (1:3000, Proteintech, HRP-66009, China), and anti-mouse/rabbit secondary antibodies (1:3000, Proteintech, SA00001-1, SA00001-2 China).

Techniques: Expressing, Two Tailed Test, Western Blot