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: Pain

Article Title: Kidney and brain-expressed protein upregulation in the anterior cingulate cortex mediates chronic post-thoracotomy pain by the phospho-protein kinase Mζ/glutamate receptor 1 signaling pathway and neuroinflammation in male rats

doi: 10.1097/j.pain.0000000000003849

Figure Lengend Snippet: GluR1 and neuroinflammation in ACC neurons are involved in CPTP. (A-F) Representative blots and columnar statistical charts show GluR1, TNF-α, and IL-1β levels in contralateral ACC at different time points in Thoracotomy pain (A-C) or Thoracotomy no pain group (D-F), * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 vs Sham group, the data were analyzed by 1-way ANOVA followed by Tukey multiple comparisons test, n = 5∼7 rats in each group. (G-I) The photographs of the double immunofluorescence staining show that GluR1, TNF-α, and IL-1β are only colocalized with neuron markers (NeuN) but not with astrocyte marker (GFAP) and microglia marker (Iba1) in the Sham and CPTP groups. The white arrowheads indicate the co-immunostaining: scale bar, 100 μm. ACC, anterior cingulate gyrus; ANOVA, analysis of variance; CPTP, chronic post-thoracotomy pain; GFAP, glial fibrillary acidic protein; GluR1, glutamate receptor 1; Iba1, ionized calcium-binding adapter molecule 1; IL-1β, interleukin-1β; NeuN, neuron-specific nuclear protein; TNF-α, tumor necrosis factor-α.

Article Snippet: After using the Pierce Bicinchoninic Acid Assay kit to measure the protein concentration, 30 μg protein was separated with SDS-PAGE gels (Bio-Rad, Hercules, CA) and immunoblotted with antibodies against KIBRA (bs-11570R; Bioss, Woburn, MA), GluR1 (67642-1-lg; Proteintech, Rosemont, IL), p -PKMζ (AF3404; Affinity, Cincinnati, OH), PKMζ (sc-17781; Santa Cruz, Dallas, TX), β-actin (EM21002; HUABIO, Hangzhou, China), tumor necrosis factor-α (TNF-α, BS-1857; Bioworld, Dublin, OH), and interleukin-1β (IL-1β, ab9722; Abcam, Cambridge, United Kingdom).

Techniques: Double Immunofluorescence Staining, Marker, Immunostaining, Binding Assay

Journal: Pain

Article Title: Kidney and brain-expressed protein upregulation in the anterior cingulate cortex mediates chronic post-thoracotomy pain by the phospho-protein kinase Mζ/glutamate receptor 1 signaling pathway and neuroinflammation in male rats

doi: 10.1097/j.pain.0000000000003849

Figure Lengend Snippet: Knockdown of KIBRA in contralateral ACC prevents the upregulation of KIBRA, p -PKMζ/GluR1 signaling pathways, and neuroinflammation in CPTP rats. (A-F) Representative blots and columnar statistical charts show KIBRA, p -PKMζ, PKMζ, TNF-α, and IL-1β levels in ACC analyzed 21 days after the sham operation, thoracotomy, or KIBRA − + thoracotomy. The data were analyzed by 1-way ANOVA followed by Tukey multiple comparisons test, n = 4∼8 rats in each group. ACC, anterior cingulate gyrus; CPTP, chronic post-thoracotomy pain; GluR1, glutamate receptor 1; IL-1β, interleukin-1β; KIBRA, kidney and brain-expressed protein; PKMζ, protein kinase Mζ; p -PKMζ, phosphorylated protein kinase Mζ; TNF-α, tumor necrosis factor-α.

Article Snippet: After using the Pierce Bicinchoninic Acid Assay kit to measure the protein concentration, 30 μg protein was separated with SDS-PAGE gels (Bio-Rad, Hercules, CA) and immunoblotted with antibodies against KIBRA (bs-11570R; Bioss, Woburn, MA), GluR1 (67642-1-lg; Proteintech, Rosemont, IL), p -PKMζ (AF3404; Affinity, Cincinnati, OH), PKMζ (sc-17781; Santa Cruz, Dallas, TX), β-actin (EM21002; HUABIO, Hangzhou, China), tumor necrosis factor-α (TNF-α, BS-1857; Bioworld, Dublin, OH), and interleukin-1β (IL-1β, ab9722; Abcam, Cambridge, United Kingdom).

Techniques: Knockdown, Protein-Protein interactions

Journal: Pain

Article Title: Kidney and brain-expressed protein upregulation in the anterior cingulate cortex mediates chronic post-thoracotomy pain by the phospho-protein kinase Mζ/glutamate receptor 1 signaling pathway and neuroinflammation in male rats

doi: 10.1097/j.pain.0000000000003849

Figure Lengend Snippet: Overexpression of KIBRA in ACC causes allodynia and activates p -PKMζ/GluR1 signaling pathways and neuroinflammation. (A) Experiment designs are shown. (B) Mechanical hyperalgesia ratio on 7, 14, and 21 days after thoracotomy or KIBRA overexpression. **** P < 0.0001 vs Thoracotomy pain group; the data were analyzed by Fisher exact test. (C and D) The threshold% and mechanical pain threshold at different time points for pain rats after injection of rAAV-CMV-wwc1-3xFLAG-WPREs or thoracotomy, * P < 0.05, *** P < 0.001, and **** P < 0.0001 vs Sham group, &&&& P < 0.00001 vs KIBRA + pain group, the data were analyzed by 2-way ANOVA followed by Tukey multiple comparisons test, n = 8 in Sham, n = 13 in Thoracotomy pain, n = 12 in KIBRA + pain, n = 8 in KIBRA + no pain group. (E-J) Representative blots and columnar statistical charts show KIBRA, p -PKMζ, PKMζ, GluR1, TNF-α, and IL-1β levels in ACC analyzed 21 days in Sham, KIBRA + pain, and KIBRA + no pain rats. The data were analyzed by 1-way ANOVA followed by Tukey multiple comparisons test, n = 4 in each group. ACC, anterior cingulate gyrus; ANOVA, analysis of variance; GluR1, glutamate receptor 1; IL-1β, interleukin-1β; KIBRA, kidney and brain-expressed protein; PKMζ, protein kinase Mζ; p -PKMζ, phosphorylated protein kinase Mζ; TNF-α, tumor necrosis factor-α.

Article Snippet: After using the Pierce Bicinchoninic Acid Assay kit to measure the protein concentration, 30 μg protein was separated with SDS-PAGE gels (Bio-Rad, Hercules, CA) and immunoblotted with antibodies against KIBRA (bs-11570R; Bioss, Woburn, MA), GluR1 (67642-1-lg; Proteintech, Rosemont, IL), p -PKMζ (AF3404; Affinity, Cincinnati, OH), PKMζ (sc-17781; Santa Cruz, Dallas, TX), β-actin (EM21002; HUABIO, Hangzhou, China), tumor necrosis factor-α (TNF-α, BS-1857; Bioworld, Dublin, OH), and interleukin-1β (IL-1β, ab9722; Abcam, Cambridge, United Kingdom).

Techniques: Over Expression, Protein-Protein interactions, Injection

Journal: Pain

Article Title: Kidney and brain-expressed protein upregulation in the anterior cingulate cortex mediates chronic post-thoracotomy pain by the phospho-protein kinase Mζ/glutamate receptor 1 signaling pathway and neuroinflammation in male rats

doi: 10.1097/j.pain.0000000000003849

Figure Lengend Snippet: ACC overexpression KIBRA combined with thoracotomy induced CPTP in all rats. (A) Experiment designs are shown. (B) Incidence of pain on 7, 14, and 21 days after thoracotomy in KIBRA overexpression rats. ** P < 0.01 vs Thoracotomy pain group; the data were analyzed by Fisher exact test. (C and D) The threshold% and mechanical pain threshold at different time points in KIBRA + no pain + Thoracotomy pain, Thoracotomy pain, or Sham group, **** P < 0.0001 vs Sham group, & P < 0.05 vs Thoracotomy pain group. The data were analyzed by 2-way ANOVA followed by Tukey multiple comparisons test, n = 8 in Sham, n = 13 in Thoracotomy pain, n = 14 in KIBRA + no pain + Thoracotomy pain group. (E-J) Representative blots and columnar statistical charts show KIBRA, p -PKMζ, PKMζ, GluR1, TNF-α, and IL-1β levels in ACC analyzed on POD21 in Sham, Thoracotomy pain, or KIBRA + no pain + Thoracotomy pain rats, n = 4 rats in each group. The data were analyzed by 1-way ANOVA followed by Tukey post hoc test. ACC, anterior cingulate gyrus; ANOVA, analysis of variance; CPTP, chronic post-thoracotomy pain; GluR1, glutamate receptor 1; IL-1β, interleukin-1β; KIBRA, kidney and brain-expressed protein; PKMζ, protein kinase Mζ; p -PKMζ, phosphorylated protein kinase Mζ; TNF-α, tumor necrosis factor-α.

Article Snippet: After using the Pierce Bicinchoninic Acid Assay kit to measure the protein concentration, 30 μg protein was separated with SDS-PAGE gels (Bio-Rad, Hercules, CA) and immunoblotted with antibodies against KIBRA (bs-11570R; Bioss, Woburn, MA), GluR1 (67642-1-lg; Proteintech, Rosemont, IL), p -PKMζ (AF3404; Affinity, Cincinnati, OH), PKMζ (sc-17781; Santa Cruz, Dallas, TX), β-actin (EM21002; HUABIO, Hangzhou, China), tumor necrosis factor-α (TNF-α, BS-1857; Bioworld, Dublin, OH), and interleukin-1β (IL-1β, ab9722; Abcam, Cambridge, United Kingdom).

Techniques: Over Expression

Journal: bioRxiv

Article Title: Granulocyte colony-stimulating factor acts through calcium-permeable AMPA receptors to potentiate cocaine reward

doi: 10.64898/2026.01.30.702629

Figure Lengend Snippet: A.Percent change PSD95 expression from saline (control) in mice treated with G-CSF (50 ug/kg), cocaine (7.5 mg/kg), or G-CSF+cocaine (50 ug/kg + 7.5 mg/kg). B . Percent change GluR1 expression from saline (control) in mice treated with G-CSF, cocaine, or G-CSF+cocaine. C . Percent change GluR2 expression from saline (control) in mice treated with G-CSF, cocaine, or G-CSF+cocaine. D . Example PSD95 blot. E . Example GluR1 blot. F . Example GluR2 blot. * p < 0.05.

Article Snippet: Proteins were transferred to a PVDF membrane and blocked with Intercept blocking buffer (LiCor) before incubation with GluR1 (1:1000, Cell Signaling Technology #13185), PSD95 (1:1000, Cell Signaling Technology #36233), or GluR2 (1:1000, Cell Signaling Technology #13607), and actin (1:5000, MP Biomedicals #691001) antibodies overnight at 4°C.

Techniques: Expressing, Saline, Control