Journal: Pediatric Research

Article Title: Spatiotemporal expression pattern of dyslexia susceptibility 1 candidate 1 (DYX1C1) during rat cerebral cortex development

doi: 10.1038/s41390-025-03920-6

Figure Lengend Snippet: a Representative images of double immunostaining of DYX1C1 (green) and a neuronal marker expressed during development DCx (red). Arrowheads in the high magnification inset show DYX1C1- and DCx-double positive cells. b DYX1C1 (green) and a neuronal marker during development, β3-tubulin (Tuj1, red). Arrowhead shows DYX1C1-positive cells co-expressing Tuj1. c DYX1C1 (green) and a neuronal differentiation factor neuroD2 (red) expression. Arrowheads show DYX1C1 and neuroD2-double positive cells. d Double immunostaining of DYX1C1 (green) and a radial glial marker nestin (red). ( a – d n = 6 ~ 8 embryos/stage) The lower panels display high-resolution images taken by a Zeiss LSM 780 confocal microscope, facilitating visualization of protein co-localization patterns. Scale bar = 500 μm (upper panels), 50 μm (lower panels), 20 μm (insets). nc neocortex, lv lateral ventricle, pia pia mater, CP cortical plate, MZ marginal zone, PCZ primitive cortical zone.

Article Snippet: Primary antibodies for fluorescence immunostaining, including mouse monoclonal anti-reelin Ab (1:1,000, CR-50) for identification of CR cells, mouse monoclonal anti-doublecortin (DCx) Ab (1:100, sc-271390; Santa Cruz Biotechnology, Santa Cruz, CA), mouse monoclonal anti-β3 tubulin Ab (1:100, Tuj1, sc-51670; Santa Cruz Biotechnology) for identification of neurons during development, mouse monoclonal anti-neuroD2 Ab (1:200, sc-365896; Santa Cruz Biotechnology) for detection of terminally differentiating neurons, mouse monoclonal anti-nestin Ab (1:1,000, ab22035; Abcam, Cambridge, UK) for identification of radial glial cells, and mouse monoclonal anti-ADP-ribosylation factor-like protein 13B (Arl13b) (1:1,000, 75-287; Antibodies Inc., Davis, CA) for visualization of primary cilia, were incubated with sections overnight at 4 °C.

Techniques: Double Immunostaining, Marker, Expressing, Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Molecular Insights of Neuroprotective Effect of Cornulaca monacantha Extract Against LPS-Induced Neuroinflammation Supported by Metabolic Profiling and Protein Interaction Analysis

doi: 10.3390/ijms27052263

Figure Lengend Snippet: Effect of C. monacantha methanolic extract on Neuro-2a cell viability. The tested concentrations of the methanolic extract ranged from 31.25 to 1000 µg/mL. Data are expressed as percentage of the untreated control (set at 100%) and presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc multiple-comparison test ( p < 0.05). a: significant vs. control; b: significant vs. 31.25 µg/mL; c: significant vs. 62.5 µg/mL; d: significant vs. 125 µg/mL.

Article Snippet: In this study, the Neuro-2a (Cat. # ABC-TC0819) mouse neuroblastoma cell line (AcceGen Biotech, Fairfield, NJ, USA) was used.

Techniques: Control, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Molecular Insights of Neuroprotective Effect of Cornulaca monacantha Extract Against LPS-Induced Neuroinflammation Supported by Metabolic Profiling and Protein Interaction Analysis

doi: 10.3390/ijms27052263

Figure Lengend Snippet: Effect of C. monacantha extract on LPS-induced neuroinflammation in Neuro-2a cells. ( a ) IL-1β and ( b ) MCP-1. Data are expressed as mean ± SD. * p < 0.05. LPS: lipopolysaccharide; IL-1β: interleukin-1β; MCP-1: monocyte chemoattractant protein-1.

Article Snippet: In this study, the Neuro-2a (Cat. # ABC-TC0819) mouse neuroblastoma cell line (AcceGen Biotech, Fairfield, NJ, USA) was used.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Molecular Insights of Neuroprotective Effect of Cornulaca monacantha Extract Against LPS-Induced Neuroinflammation Supported by Metabolic Profiling and Protein Interaction Analysis

doi: 10.3390/ijms27052263

Figure Lengend Snippet: Effect of C. monacantha extract on the mRNA expression of the Nrf2 pathway and related genes in Neuro-2a cells. ( a ) Nrf2, ( b ) NF-κB, ( c ) Hmox1, and ( d ) NQO-1. Data are expressed as mean ± SD. * p < 0.05. LPS: lipopolysaccharide; Nrf2: nuclear factor erythroid 2–related factor 2; NF-κB: nuclear factor kappa B; Hmox1: heme oxygenase 1; NQO-1: NAD(P)H quinone dehydrogenase 1.

Article Snippet: In this study, the Neuro-2a (Cat. # ABC-TC0819) mouse neuroblastoma cell line (AcceGen Biotech, Fairfield, NJ, USA) was used.

Techniques: Expressing

Journal: Cell communication and signaling : CCS

Article Title: Assessment of technical and clinical utility of a bead-based flow cytometry platform for multiparametric phenotyping of CNS-derived extracellular vesicles.

doi: 10.1186/s12964-023-01308-9

Figure Lengend Snippet: Fig. 1 Graphic illustration of the different combinations of sample material and EV isolation protocols applied to the EV Neuro assay. For further details, see text. Abbreviations: HA: primary human astrocytes; GB: glioblastoma; HC: healthy control; MS: multiple sclerosis; MAD: mild cognitive impairment/ Alzheimer’s disease/depression; SEC: size-exclusion chromatography; UC: ultracentrifugation

Article Snippet: MACSPlex EV Kit Neuro, human, reagents were kindly provided by Miltenyi Biotec (Bergisch Gladbach).

Techniques: Isolation, Control, Size-exclusion Chromatography

Journal: Cell communication and signaling : CCS

Article Title: Assessment of technical and clinical utility of a bead-based flow cytometry platform for multiparametric phenotyping of CNS-derived extracellular vesicles.

doi: 10.1186/s12964-023-01308-9

Figure Lengend Snippet: Fig. 2 Characterization of EVs derived from glioblastoma cells and astrocytes. a Size distribution of EVs separated from cell culture supernatant of LN18, LN229, and NCH82 glioblastoma cell lines as well as from primary human astrocytes (HA). b Western blot analysis of CD9, CD63, CD81, Syntenin (EV markers) and Calnexin (non-EV marker) in EVs separated from cell culture supernatant of glioblastoma cells and primary astrocytes. c EV Neuro median signal intensities for LN18, LN229, NCH82, and HA EVs. d Relative signal intensities of values for cell derived EVs shown in (c). Relative signals are calculated by dividing the signal intensity of a target by the sum of intensities revealed from all markers (total intensity) and presented in percentages. Bars reflect representative marker profiles (n = 1–3 biological replicates). Arrow heads indicate markers that appear elevated in cell lines compared to primary astrocytes

Article Snippet: MACSPlex EV Kit Neuro, human, reagents were kindly provided by Miltenyi Biotec (Bergisch Gladbach).

Techniques: Derivative Assay, Cell Culture, Western Blot, Marker

Journal: Cell communication and signaling : CCS

Article Title: Assessment of technical and clinical utility of a bead-based flow cytometry platform for multiparametric phenotyping of CNS-derived extracellular vesicles.

doi: 10.1186/s12964-023-01308-9

Figure Lengend Snippet: Fig. 3 Titration of EVs derived from glioblastoma cells. EV Neuro median signal intensities for increasing total numbers of NCH82 EVs (a) and LN18 EVs (b) as determined by NTA particle count. The number of 2.8 × 107 particles (turquoise) represents the input of 120 µl of pre-cleared cell culture supernatant (10,000 × g) without prior EV enrichment. Other particle inputs (blue and red) were achieved by EV enrichment via dUC. Bars reflect marker profiles of one experiment each

Article Snippet: MACSPlex EV Kit Neuro, human, reagents were kindly provided by Miltenyi Biotec (Bergisch Gladbach).

Techniques: Titration, Derivative Assay, Cell Culture, Marker

Journal: Cell communication and signaling : CCS

Article Title: Assessment of technical and clinical utility of a bead-based flow cytometry platform for multiparametric phenotyping of CNS-derived extracellular vesicles.

doi: 10.1186/s12964-023-01308-9

Figure Lengend Snippet: Fig. 4 Profiling of serum EVs from GB patients and healthy controls. a Relative EV Neuro signal intensities for EVs separated from the serum of GB patients or HC by SEC followed by UC calculated as signal of target divided by the total signal of all markers (in %). Bars represent mean values and error bars indicate the 95% confidence interval. Asterisks mark statistically significant differences between GB and HC. Only significant alterations of targets that were consistently detected above background in all individuals are marked. *=p < .05, **=p < .01, ***=p < .001. (b) tSNE on data from (a) stratified by condition (color). c Heatmap visualization of selected targets from (a) including hierarchical clustering for targets as well as subjects

Article Snippet: MACSPlex EV Kit Neuro, human, reagents were kindly provided by Miltenyi Biotec (Bergisch Gladbach).

Techniques:

Journal: Cell communication and signaling : CCS

Article Title: Assessment of technical and clinical utility of a bead-based flow cytometry platform for multiparametric phenotyping of CNS-derived extracellular vesicles.

doi: 10.1186/s12964-023-01308-9

Figure Lengend Snippet: Fig. 6 Comparison of plasma EVs from MS patients in a relapse versus in a stable disease phase. Relative EV Neuro signal intensities as signal of target divided by the total signal of all markers (in %) for immuno-affinity captured CD63+EVs (a) and CD81+EVs (b). Bars represent mean values and error bars indicate the 95% confidence interval. Asterisks mark statistically significant differences between stable phase and disease relapse. Only significant alterations of targets that were consistently detected above background in all individuals are marked. *=p < .05, **=p < .01

Article Snippet: MACSPlex EV Kit Neuro, human, reagents were kindly provided by Miltenyi Biotec (Bergisch Gladbach).

Techniques: Comparison, Clinical Proteomics

Journal: Cell communication and signaling : CCS

Article Title: Assessment of technical and clinical utility of a bead-based flow cytometry platform for multiparametric phenotyping of CNS-derived extracellular vesicles.

doi: 10.1186/s12964-023-01308-9

Figure Lengend Snippet: Fig. 7 Phenotyping of plasma EVs from MAD patients and comparison of all plasma samples. a Relative EV Neuro signal intensities for EVs separated from the plasma of MAD patients by immuno-affinity capture (anti-CD81) and magnetic separation. Bars represent mean values and error bars indicate the 95% confidence intervals (n = 8 patients). b tSNE of relative signal intensities from all plasma-derived EV samples analyzed (MS, MAD, HC) stratified by condition (color) and isolation (shape). c Heatmap visualization of relative signals for selected targets from all plasma samples analyzed including hierarchical clustering for targets as well as subjects (BCL = lab of blood collection)

Article Snippet: MACSPlex EV Kit Neuro, human, reagents were kindly provided by Miltenyi Biotec (Bergisch Gladbach).

Techniques: Clinical Proteomics, Comparison, Derivative Assay, Isolation

Journal: Journal of Neurosurgery

Article Title: Neuron-derived orphan receptor 1 transduces survival signals in neuronal cells in response to hypoxia-induced apoptotic insults

doi: 10.3171/2015.6.jns1535

Figure Lengend Snippet: Fig. 3. Roles of CREB in hypoxia-induced NOR-1 mRNA expression. Neuro-2a cells were exposed to OGD for 1, 3, and 6 hours. Levels of nuclear CREB were immunodetected (A, upper panel). Amounts of PCNA were measured as the internal standard (lower panel). These protein bands were quantified and statistically analyzed (B). Neuro-2a cells were exposed to CREB siRNA (siRNA) for 48 hours. Control cells received scrambled siRNA. Levels of nuclear CREB were immunodetected (C, upper panel). PCNA was measured as the internal standard. These protein bands were quantified and statistically analyzed (lower panel). Neuro-2a cells were treated with OGD, CREB siRNA (siRNA), and a combination of siRNA and OGD for 6 hours. Levels of NOR-1 mRNA were analyzed using a quantitative PCR (D). Each value in the bar graph represents the mean ± SEM (n = 6). *#Values significantly (p < 0.05) differed from those of the control and OGD-treated groups, respectively.

Article Snippet: Scrambled siRNA, purchased from Santa Cruz Biotechnology, was applied to neuro-2a cells as a negative standard. animal model of tbi Male ICR mice (weighting 20–25 g) were purchased from the Animal Center of the College of Medicine, National Taiwan University.

Techniques: Expressing, Control, Real-time Polymerase Chain Reaction

Journal: Journal of Neurosurgery

Article Title: Neuron-derived orphan receptor 1 transduces survival signals in neuronal cells in response to hypoxia-induced apoptotic insults

doi: 10.3171/2015.6.jns1535

Figure Lengend Snippet: Fig. 4. Roles of cIAP2 in hypoxia-induced neuronal insults. Neuro-2a cells were exposed to OGD for 1, 3, and 6 hours. Levels of cIAP2 were immunodetected (A, upper panel). Amounts of b-actin were detected as the internal standard (lower panel). These protein bands were quantified and statistically analyzed (B). Levels of cIAP2 mRNA were analyzed using quantitative PCR analy- ses (C). Neuro-2a cells were exposed to cIAP2 siRNA (siRNA) for 48 hours and then treated with OGD, and a combination of OGD and siRNA for 6 hours. Scrambled siRNA was administered to control cells as the negative control. Cell viability was determined by a colorimetric method (D). Each value represents the mean ± SEM (n = 6). Each value represents the mean ± SEM (n = 6). *#Values significantly (p < 0.05) differed from those of the control and OGD-treated groups, respectively.

Article Snippet: Scrambled siRNA, purchased from Santa Cruz Biotechnology, was applied to neuro-2a cells as a negative standard. animal model of tbi Male ICR mice (weighting 20–25 g) were purchased from the Animal Center of the College of Medicine, National Taiwan University.

Techniques: Real-time Polymerase Chain Reaction, Control, Negative Control

Journal: Journal of Neurosurgery

Article Title: Neuron-derived orphan receptor 1 transduces survival signals in neuronal cells in response to hypoxia-induced apoptotic insults

doi: 10.3171/2015.6.jns1535

Figure Lengend Snippet: Fig. 5. Participation of NOR-1 in regulation of cIAP2 mRNA expression. NOR-1 siRNA was administered to neuro-2a cells for 48 hours. Control cells received scrambled siRNA as the negative control. Levels of NOR- 1 were immunodetected (A, upper panel). b-Actin was quantified as the internal standard (lower panel). These protein bands were quantified and statistically analyzed (B). Neuro-2a cells were exposed to OGD, NOR- 1 siRNA (siRNA), and a combination of OGD and siRNA for 6 hours. NOR-1 mRNA was analyzed using quantitative PCR (C). Each value represents the mean ± SEM (n = 6). *#Values significantly (p < 0.05) differed from those of the control and OGD-treated groups, respectively.

Article Snippet: Scrambled siRNA, purchased from Santa Cruz Biotechnology, was applied to neuro-2a cells as a negative standard. animal model of tbi Male ICR mice (weighting 20–25 g) were purchased from the Animal Center of the College of Medicine, National Taiwan University.

Techniques: Expressing, Control, Negative Control, Real-time Polymerase Chain Reaction

Journal: Journal of Neurosurgery

Article Title: Neuron-derived orphan receptor 1 transduces survival signals in neuronal cells in response to hypoxia-induced apoptotic insults

doi: 10.3171/2015.6.jns1535

Figure Lengend Snippet: Fig. 6. Roles of NOR-1 in OGD-induced apoptotic insults to neuronal cells. Neuro-2a cells were exposed to OGD, NOR-1 siRNA, and a combination of OGD and siRNA for 6 hours. Control cells received Scrambled siRNA as the negative control. Cell viability was assayed with a colorimetric method (A). Caspase-3 activity was analyzed using a fluorometric substrate assay (B). DNA fragmentation was quantified using an ELISA method (C). Apoptotic cells were quantified using a flow cytometer (D). Each value represents the mean ± SEM (n = 6). *#Values significantly (p < 0.05) differed from those of the control and OGD-treated groups, respectively.

Article Snippet: Scrambled siRNA, purchased from Santa Cruz Biotechnology, was applied to neuro-2a cells as a negative standard. animal model of tbi Male ICR mice (weighting 20–25 g) were purchased from the Animal Center of the College of Medicine, National Taiwan University.

Techniques: Control, Negative Control, Activity Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry