Journal: Alzheimer's Research & Therapy

Article Title: IRF1 ameliorates synaptic dysfunction through the modulation of O-GlcNAcylation on GluN1 subunit of NMDAR

doi: 10.1186/s13195-025-01857-w

Figure Lengend Snippet: IRF-1 modulates the O-GlcNAcylation of GluN1. ( A ) Cortical brain homogenates were collected from 3xTg-AD mice injected with AAV-IRF-1 in the hippocampus to assess NMDARs expression through western blotting analysis ( n = 3). ( B-D ) The O-GlcNAcylation levels of immunoprecipitated GluN1 (B), GluN2A (C) or GluN2B (D) from brain homogenates of 3xTg-AD mice injected with AAV-IRF-1 were analyzed using western blots with anti-RL2, anti-GluN1, anti-GluN2A or anti-GluN2B antibody. The relative O-GlcNAcylation levels (RL2) of immunoprecipitated GluN1, GluN2A or GluN2B were quantified after normalization with GluN1, GluN2A and GluN2B ( n = 3). ( E-H ) Mass Spectrometry analysis was performed on GluN1 immunoprecipitated from brain homogenates of IRF-1 knockout and their littermate control mice. All data are presented as mean ± S.E.M. * p < 0.05 vs. pcDNA, AAV-NC or WT, two-tailed student’s t test or two-way ANOVA with Šídák’s multiple comparisons test

Article Snippet: Primary neuronal cells were infected with shIRF-1 adenovirus for 48 h. The cells were then incubated with medium-diluted rabbit-derived antibodies against GluN1 (Cell Signaling Technology), GluN2A (Cell Signaling Technology), and GluN2B (Proteintech) for 20 min at 37 °C.

Techniques: Injection, Expressing, Western Blot, Immunoprecipitation, Mass Spectrometry, Knock-Out, Control, Two Tailed Test

Journal: Alzheimer's Research & Therapy

Article Title: IRF1 ameliorates synaptic dysfunction through the modulation of O-GlcNAcylation on GluN1 subunit of NMDAR

doi: 10.1186/s13195-025-01857-w

Figure Lengend Snippet: IRF-1 reduces the internalization of GluN1 and increases its surface expression. ( A-B ) Immunocytochemical images (A) and quantitative analysis (B) of internalized and total GluN1, GluN2A, and GluN2B in primary neuronal cells infected with NC and shIRF-1 adenovirus ( n = 12). ( C-F ) Immunoblots displaying surface and total levels of NMDAR subunits in primary neuronal cells infected with IRF-1 adenovirus (C-D) and in cortical slices of 3xTg-AD mice injected with AAV-IRF-1 in the hippocampus (E-F) ( n = 3). ( G-I ) Representative images of dendritic spines in hippocampal pyramidal neurons and quantification of dendritic spines. Dendritic spine density per 10 μm in hippocampal pyramidal neurons of 3xTg-AD mice injected with AAV-IRF-1 and AAV-NC mice, determined by Golgi staining and percentages of stubby, mushroom, thin, or filopodium spines were quantified ( n = 10). All data are presented as mean ± S.E.M. * p < 0.05; ** p < 0.01; *** p < 0.001 vs. NC, AAV-NC, two-tailed student’s t test or two-way ANOVA with Šídák’s multiple comparisons test

Article Snippet: Primary neuronal cells were infected with shIRF-1 adenovirus for 48 h. The cells were then incubated with medium-diluted rabbit-derived antibodies against GluN1 (Cell Signaling Technology), GluN2A (Cell Signaling Technology), and GluN2B (Proteintech) for 20 min at 37 °C.

Techniques: Expressing, Infection, Western Blot, Injection, Staining, Two Tailed Test