Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: A Outline of study time-course and sample processing for EV isolation. Antemortem plasma samples with postmortem pathological confirmation of neurological diagnoses were processed to isolate cell-specific EVs using our mTENPO microfluidic platform, alongside plasma protein biomarkers using commercial digital ELISA, for patients with LBD ( n = 30), AD ( n = 31), AD/LBD ( n = 30), AD/ALB ( n = 19), and controls ( n = 27). B The mTENPO platform, illustrating the external magnet, inlet reservoir, outlet ports, and tubing connections to syringe pumps. Syringes are connected to the waste outlet for blocking, washing, and sample addition steps, and then replaced and switched to the lysate outlet before captured EVs are lysed on-chip. The inset shows a photo of the mTENPO chip with a quarter for scale. C Schematic of operation of the mTENPO platform for cell-specific EV isolation using antibody-labeled magnetic nanoparticles (MNPs) for GluR2+ (top) and GLAST+ (bottom) EV pulldowns. D Scanning electron microscopy (SEM) images of GluR2+ EVs immobilized on the edges of pores of the mTENPO device’s surface. E SEM images of GLAST+ EVs immobilized on the edges of pores of the mTENPO device’s surface. F Representative cropped western blot images showing protein expression of GluR2, GLAST, and EV-associated marker TSG101 using mTENPO-isolated GluR2+ or GLAST + EV lysates from n = 2 human plasma samples. Full-length western blot images are shown in Supplementary Fig. .

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Isolation, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Blocking Assay, Labeling, Electron Microscopy, Western Blot, Expressing, Marker

Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: A Heatmap of z-score of log 2 (expression) for biomarkers with Benjamini-Hochberg FDR-corrected P value < 0.1. Subjects (columns) are hierarchically clustered within cohort and biomarkers within each compartment (rows) are sorted by descending fold-change. B Volcano plot demonstrating differential expression of GluR2+ EV miRNAs, GLAST + EV miRNAs, and plasma proteins. C Venn diagram showing overlap in FDR P value significant miRNAs ( P value < 0.1) between GluR2+ EVs and GLAST+ EVs. D Top 30 biomarkers in all compartments ranked by descending area under the curve (AUC). Error bars represent standard error from bootstrapping 10x.

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Expressing, Quantitative Proteomics, Clinical Proteomics

Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: GO and KEGG pathway analyses were performed on differentially expressed miRNAs using DIANA miRPath v4.0 using the TarBase v8.0 database. FDR P values for identified GO terms and KEGG pathways were calculated using a one-sided Fisher’s exact test and considered significant at P value < 0.05. The top 10 (ranked by number of target genes) terms within each of the three GO categories (BP, CC, MF) and top 10 (ranked by number of target genes) KEGG pathways were identified for each pulldown. A Top 10 terms within each GO category for GluR2+ EV miRNAs. B Top 10 KEGG pathways for GluR2+ EV miRNAs. C Top 10 terms within each GO category for GLAST + EV miRNAs. D Top 10 KEGG pathways for GLAST + EV miRNAs. In all panels, each bar is labeled to the right with the number of differentially expressed miRNAs associated with the given GO term or KEGG pathway.

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Labeling

Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: A Heatmap of z-score of log 2 (expression) for LASSO-selected biomarkers. Subjects (columns) are hierarchically clustered within cohort and biomarkers within each compartment (rows) are sorted by descending AUC. B Kendall correlation staircase plots identifying the extent to which biomarker information was correlated between the LASSO-selected GluR2+ EV, GLAST + EV, and protein biomarkers. Biomarkers are sorted within compartments by AUC. The inset shows the correlation distribution of Kendall’s τ, where the dotted line represents the median count. C LASSO panel accuracy versus panel size for classifying LBD versus AD, shown in blue; accuracy is assessed through tenfold cross-validation, with error bars representing standard error from 5 repeats of panel training on the LBD vs AD patient groups. Average accuracy and standard error for control experiments performed by scrambling patient labels 10x are shown in orange. D LASSO panel AUC versus panel size for classifying LBD versus AD, shown in blue with error bars as described in ( C ). Average AUC and standard error for the same control experiments described in ( C ) are shown in orange. E AUCs for the 15-marker LASSO panel and individual LASSO biomarkers, sorted by descending AUC. Error bars represent 95% confidence intervals, calculated from 5x repeats of tenfold cross-validation for the 15-marker panel or from bootstrapping 10x for individual markers.

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Expressing, Biomarker Discovery, Control, Marker

Journal: Neuron

Article Title: Regulation of Thalamic and Cortical Network Synchrony by Scn8a

doi: 10.1016/j.neuron.2017.01.031

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Monoclonal mouse anti-GluR2 , Neuromab , Cat# 75-002; RRID: AB_2232661.

Techniques: Plasmid Preparation, Virus, Recombinant, Avidin-Biotin Assay, Software, Imaging

Journal: Neural Regeneration Research

Article Title: Sequential expression of cyclooxygenase-2, glutamate receptor-2, and platelet activating factor receptor in rat hippocampal neurons after fluid percussion injury

doi: 10.4103/1673-5374.133151

Figure Lengend Snippet: PAFR (381 bp), COX-2 (287 bp), and GluR2 (352 bp) mRNA expression detected by RT-PCR in hippocampal neurons at different injury time points. M: Marker; C: control group; 4 h, 8 h, 12 h, 24 h, and 48 h: post-injury 4, 8, 12, 24, and 48 h groups; h: hours. Marker molecular weight is 2,000, 1,000, 750, 500, 250, and 100 bp from top to bottom. PAFR: Platelet-activating factor receptor; COX-2: cyclooxygenase-2; GluR2: glutamate receptor 2. β-Actin was used for normalization purposes.

Article Snippet: Cells were blocked with 10% rabbit serum at room temperature and incubated overnight at 37°C with the following antibodies: goat anti-PAFR, COX-2, and GluR2 polyclonal antibodies (1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA), followed by biotinylated goat anti-goat IgG (1:200; Beijing Zhongshan Biotechnology Co., Ltd.) for 30 minutes at 37°C.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Marker, Control, Molecular Weight

Journal: Neural Regeneration Research

Article Title: Sequential expression of cyclooxygenase-2, glutamate receptor-2, and platelet activating factor receptor in rat hippocampal neurons after fluid percussion injury

doi: 10.4103/1673-5374.133151

Figure Lengend Snippet: Immunohistochemical staining of COX-2, GluR2, and PAFR in hippocampal neurons 4–48 hours after injury. Arrows indicate positively stained cells for COX-2 (A), GluR2 (B), and PAFR (C) (× 200). Areas of positive hippocampal neurons in the different injury groups were compared to the control group (D). * P < 0.01, vs . control group. Data are expressed as mean ± SD of six dishes of cells for each group (one-way analysis of variance and least significant difference test). COX-2: Cyclooxygenase-2; GluR2: glutamate receptor 2; PAFR: platelet-activating factor receptor; h: hours.

Article Snippet: Cells were blocked with 10% rabbit serum at room temperature and incubated overnight at 37°C with the following antibodies: goat anti-PAFR, COX-2, and GluR2 polyclonal antibodies (1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA), followed by biotinylated goat anti-goat IgG (1:200; Beijing Zhongshan Biotechnology Co., Ltd.) for 30 minutes at 37°C.

Techniques: Immunohistochemical staining, Staining, Control

Journal: eLife

Article Title: RNG105/caprin1, an RNA granule protein for dendritic mRNA localization, is essential for long-term memory formation

doi: 10.7554/eLife.29677

Figure Lengend Snippet: ( A and B ) Immunostaining for GluR1 ( A ) and GluR2 ( B ) in cultured neurons (9 DIV) from the cerebral cortex of E17.5 Rng105 +/+ and Rng105 −/− littermates. The neurons were cultured with (+) or without (-) TTX and APV prior to the staining. GluR1 and GluR2 staining before permeabilization (green, surface proteins), after permeabilization (magenta, intracellular and residual surface proteins), and merged images (total proteins) are shown. GluR1 and GluR2 are distributed in a punctate manner both in the soma and dendrites. The insets show magnified images of boxed areas. Arrowheads denote representative GluR1 and GluR2 puncta which were stained both before and after permeabilization (white), only before permeabilization (yellow) and only after permeabilization (blue). Scale bars, 10 µm. ( C and D ) Quantitative analysis of GluR1 and GluR2 surface expression in dendrites. C, the number of surface GluR1 puncta in dendrites normalized by the number of total GluR1 puncta (left), and fluorescence intensity of surface GluR1 puncta in dendrites normalized by GluR1 fluorescence intensity after permeabilization and in the soma (right). D, the same quantification for GluR2. Data are represented as the mean ± s.e.m. In C, n = 31 ( Rng105 +/+ , −), 35 ( Rng105 +/+ , +), 34 ( Rng105 −/− , −), and 33 ( Rng105 −/− , +) neurons from 4 experiments. In D, n = 39 ( Rng105 +/+ , −), 40 ( Rng105 +/+ , +), 39 ( Rng105 −/− , −), and 38 ( Rng105 −/− , +) neurons from 4 experiments. ***p<0.005, ****p<0.001 using two-way ANOVA followed by post-hoc Student's t-test. See also .

Article Snippet: The total lysate and biotinylated eluate were analyzed by western blotting with the anti-GluR1 (1:50, PC246, Merck Millipore), and anti-GluR2 (1:1000, MAB397, Merck Millipore) antibodies.

Techniques: Immunostaining, Cell Culture, Staining, Expressing, Fluorescence

Journal: eLife

Article Title: RNG105/caprin1, an RNA granule protein for dendritic mRNA localization, is essential for long-term memory formation

doi: 10.7554/eLife.29677

Figure Lengend Snippet: ( A ) Total cell lysates of surface-biotinylated primary cultured neurons (9 DIV) from E17.5 wild-type mouse cerebral cortex (total), and avidin agarose beads-bound fractions of the lysates (surface), were immunoblotted with the anti-GluR1 antibody. Control neurons were mock-treated without biotin. Arrow and arrowhead indicate biotin-labeled surface GluR1 and non-labeled intracellular GluR1, respectively. Lanes were cut and moved horizontally in the same membrane. ( B and C ) TTX/APV-treated and untreated primary cultured neurons (9 DIV) from Rng105 +/+ and Rng105 −/− littermates (E17.5) were surface biotinylated and analyzed as in A. Immunoblotting for GluR1 ( B ) and GluR2 ( C ). Arrows and arrowheads indicate biotin-labeled surface GluR1/2 and non-labeled intracellular GluR1/2, respectively. In the bottom panel in B, twice the amount of samples from Rng105 −/− neurons were loaded, which showed more clearly that the ratio of surface/intracellular GluR1 was lower in Rng105 −/− neurons than in Rng105 +/+ neurons. In A−C, numbers on the left indicate molecular mass (kDa). ( D and E ) Quantitative analysis of the ratio of surface/intracellular GluR1 and GluR2 in the biotinylation assay. The intensity of upper GluR1/2 bands in the avidin beads-bound fraction (arrows in B and C) and lower GluR1/2 bands in the total lysate (arrowheads in B and C) was measured and the surface/intracellular ratio was calculated. D, GluR1; E, GluR2. Data are represented as the mean ± s.e.m. n = 9 from 3 littermates each of Rng105 +/+ and Rng105 −/− mice. *p<0.05 using two-way ANOVA followed by post-hoc Student's t-test. Attached Files.

Article Snippet: The total lysate and biotinylated eluate were analyzed by western blotting with the anti-GluR1 (1:50, PC246, Merck Millipore), and anti-GluR2 (1:1000, MAB397, Merck Millipore) antibodies.

Techniques: Cell Culture, Avidin-Biotin Assay, Labeling, Western Blot, Cell Surface Biotinylation Assay