Journal: Biomolecules

Article Title: Functional, Morphological and Molecular Changes Reveal the Mechanisms Associated with Age-Related Vestibular Loss

doi: 10.3390/biom13091429

Figure Lengend Snippet: Age-related changes in the numbers of pre- and postsynaptic synaptic elements in the vestibular sensory epithelia between young (1.5-month-old) and old (24-month-old) mice. ( A ). Age-related changes in the numbers of pre-synaptic ribbons and postsynaptic glutamate receptor patches were investigated using immunofluorescence in the striolar region of the utricular macula. Sensory epithelia were immunostained with antibodies against CTBP2 (green) to identify presynaptic ribbons and GluR2/3 (red) to identify postsynaptic glutamate receptor patches (red). ( B ). There was no significant difference in the density of presynaptic ribbons (CTBP2). ( C ). There was no significant difference in the density of postsynaptic glutamate receptor patches (GluR2/3). Non-significant differences are indicated by n.s.

Article Snippet: The first combination included mouse monoclonal (IgG1) anti-CTBP2 (BD Biosciences 612044; RRID: AB_399431, San Jose, CA, USA) and rabbit polyclonal anti-GluR2/3 (Millipore AB1506; RRID: AB_90710, Burlington, MA, USA).

Techniques: Immunofluorescence

Journal: PLoS ONE

Article Title: AMPA Receptor Regulation at the mRNA and Protein Level in Rat Primary Cortical Cultures

doi: 10.1371/journal.pone.0025350

Figure Lengend Snippet: Double labeling of rat cortical neurons at DIV5 and DIV26 (20×) with an antibody against the glial marker GFAP (green) and with antibodies against the four AMPA receptor subunits: (A, B) GluR1; (C, D) GluR2; (E, F) GluR3 and (G, H) GluR4 (red). The images were acquired from rare regions with high density of glial cells to show that the GFAP-positive cells do not express AMPA receptors. Scale bar: 50 µm.

Article Snippet: The membranes were blocked for 60 min with 3% nonfat dry milk or 2% BSA in TBS-T (Tris-buffered saline with 0.1% Tween-20, Sigma-Aldrich) and then incubated overnight at 4°C in the blocking solution with the primary antibodies corresponding to either rabbit polyclonal anti-GluR1 (1∶200, Millipore), rabbit polyclonal anti-GluR2 (1∶2500, Millipore), rabbit polyclonal anti-GluR3 (1∶500, Alomone Labs Ltd) or rabbit polyclonal anti-GluR4 (1∶100, Millipore) with mouse monoclonal anti-GAPDH (1∶2500, Sigma-Aldrich) as a loading control.

Techniques: Labeling, Marker

Journal: PLoS ONE

Article Title: AMPA Receptor Regulation at the mRNA and Protein Level in Rat Primary Cortical Cultures

doi: 10.1371/journal.pone.0025350

Figure Lengend Snippet: (A) The graphs represent the percentage of the flip isoform among the total flip + flop isoforms present during the maturation of cortical cultured neurons and in extracts of adult rat prefrontal/frontal cortex (P/FC). (B, C) Editing levels at the R/G site of the flip (B) and flop (C) variants of subunit GluR2-4 mRNAs. The results are the mean ± S.E. of 3 independent experiments, performed in independent preparations. Statistical analysis was performed by one-way ANOVA followed by the Bonferroni's multiple comparison test (*p<0.05, **p<0.01, ***p<0.001).

Article Snippet: The membranes were blocked for 60 min with 3% nonfat dry milk or 2% BSA in TBS-T (Tris-buffered saline with 0.1% Tween-20, Sigma-Aldrich) and then incubated overnight at 4°C in the blocking solution with the primary antibodies corresponding to either rabbit polyclonal anti-GluR1 (1∶200, Millipore), rabbit polyclonal anti-GluR2 (1∶2500, Millipore), rabbit polyclonal anti-GluR3 (1∶500, Alomone Labs Ltd) or rabbit polyclonal anti-GluR4 (1∶100, Millipore) with mouse monoclonal anti-GAPDH (1∶2500, Sigma-Aldrich) as a loading control.

Techniques: Cell Culture

Journal: PLoS ONE

Article Title: AMPA Receptor Regulation at the mRNA and Protein Level in Rat Primary Cortical Cultures

doi: 10.1371/journal.pone.0025350

Figure Lengend Snippet: Editing levels at the R/G site of subunits GluR2-4 for the flip and flop variants. The effects of KCl and APV/TTX treatments on the editing levels at the R/G site are shown separately for each variant. The results are expressed as the mean ± S.E. of 3 independent experiments performed in independent preparations. Statistical analysis was performed by one-way ANOVA followed by the Dunnett test (*p<0.05, **p<0.01, ***p<0.001).

Article Snippet: The membranes were blocked for 60 min with 3% nonfat dry milk or 2% BSA in TBS-T (Tris-buffered saline with 0.1% Tween-20, Sigma-Aldrich) and then incubated overnight at 4°C in the blocking solution with the primary antibodies corresponding to either rabbit polyclonal anti-GluR1 (1∶200, Millipore), rabbit polyclonal anti-GluR2 (1∶2500, Millipore), rabbit polyclonal anti-GluR3 (1∶500, Alomone Labs Ltd) or rabbit polyclonal anti-GluR4 (1∶100, Millipore) with mouse monoclonal anti-GAPDH (1∶2500, Sigma-Aldrich) as a loading control.

Techniques: Variant Assay

Journal: PLoS ONE

Article Title: AMPA Receptor Regulation at the mRNA and Protein Level in Rat Primary Cortical Cultures

doi: 10.1371/journal.pone.0025350

Figure Lengend Snippet: (A) Western blot of the four AMPA receptor subunits after 24-hour treatment with glutamate on mature cortical cultures. The results are expressed as the mean ± S.E. of 3 independent experiments performed in independent preparations and reported as percentage of non-treated control (CNT). (B) The graphs represent the percentage of the flip isoform among the total flip + flop isoforms after glutamate treatment. (C) Editing levels at the R/G site of subunits GluR2-4 for the flip and flop variants. The results are expressed as the mean ± S.E. of 3 independent experiments performed in independent preparations. Statistical analysis was performed by unpaired Student's t-tests (*p<0.05, **p<0.01, ***p<0.001).

Article Snippet: The membranes were blocked for 60 min with 3% nonfat dry milk or 2% BSA in TBS-T (Tris-buffered saline with 0.1% Tween-20, Sigma-Aldrich) and then incubated overnight at 4°C in the blocking solution with the primary antibodies corresponding to either rabbit polyclonal anti-GluR1 (1∶200, Millipore), rabbit polyclonal anti-GluR2 (1∶2500, Millipore), rabbit polyclonal anti-GluR3 (1∶500, Alomone Labs Ltd) or rabbit polyclonal anti-GluR4 (1∶100, Millipore) with mouse monoclonal anti-GAPDH (1∶2500, Sigma-Aldrich) as a loading control.

Techniques: Western Blot

Journal: iScience

Article Title: KIBRA regulates activity-induced AMPA receptor expression and synaptic plasticity in an age-dependent manner

doi: 10.1016/j.isci.2022.105623

Figure Lengend Snippet:

Article Snippet: GluA2, rabbit polyclonal , Alomone Labs , Cat# AGC-005 RRID: AB_2039881.

Techniques: Recombinant, Blocking Assay, Software