Vector Laboratories
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Journal: Neural Regeneration Research
Article Title: Protein arginine methyltransferase-6 regulates heterogeneous nuclear ribonucleoprotein-F expression and is a potential target for the treatment of neuropathic pain
doi: 10.4103/NRR.NRR-D-23-01539
Figure Lengend Snippet: PRMT6 is mainly expressed in mouse DRG nociceptive neurons. (A) In DRG neurons, β-tubulin III (green) co-localized with PRMT6 (red). (B) Astrocyte GS (green) did not co-localize with PRMT6 (red). Nuclei were stained with DAPI (blue). (C) Distribution of PRMT6 + somata: large (18.29%), small (21.95%), and medium (60.16%). (D–F) PRMT6 + neurons were stained for NF200 (green), CGRP (green), or IB4 (green), scale bars: 50 µm. Five sections per mouse from three mice per group were evaluated. CGRP: Calcitonin gene-related peptide; DAPI: 4′,6-diamidino-2-phenylindole; DRG: dorsal root ganglion; GS: glutamine synthetase; IB4: isolectin B4; NF200: neurofilament-200; PRMT6: protein arginine methyltransferase-6.
Article Snippet: The sections were blocked overnight in 5% BSA and 1% Triton X-100 in PBS and then incubated with primary antibodies against PRMT6 (rabbit, 1:200; Novus Biologicals, Littleton, CO, USA, Cat# NB100-56642, RRID: AB_838734), heterogeneous nuclear ribonucleoprotein F (hnRNP-F) (mouse,1:200; Thermo Fisher Scientific, Waltham, MA, USA, Cat# MA5-18024, RRID: AB_2539408), calcitonin gene-related peptide (CGRP; mouse, 1:200, Abcam, Cambridge, MA, USA, Cat# ab81887, RRID: AB_1658411),
Techniques: Staining
Journal: Neural Regeneration Research
Article Title: Protein arginine methyltransferase-6 regulates heterogeneous nuclear ribonucleoprotein-F expression and is a potential target for the treatment of neuropathic pain
doi: 10.4103/NRR.NRR-D-23-01539
Figure Lengend Snippet: Prmt6 –/– mice exhibit normal innervation patterns and sensory neuron numbers. (A) L3–L4 spinal cord segments were harvested from WT or Prmt6 –/– mice and immunostained for IB4 (red) and CGRP (green) to label central nociceptive terminals. The pixel density of IB4 and CGRP in the dorsal horn of the spinal cord (expressed as arbitrary units (AUs)) was quantified in ImageJ and showed no change in central innervation density in Prmt6 –/– mice. (B) Analysis of the total number of neurons in the DRG in WT and Prmt6 –/– mice. L4–L5 DRGs were harvested from WT or Prmt6 –/– mice, and three sections from each mouse were immunostained with the pan-neural marker β-tubulin III (green) and counterstained with DAPI (blue). WT and Prmt6 –/– mice exhibited similar numbers of sensory neurons. (C) Immunostaining and quantification of WT and Prmt6 –/– mouse nerve fibers. β-tubulin III was used to label all nerve fibers, IB4 to label nonpeptidergic nociceptor fibers, and CGRP to label peptidergic nociceptor fibers. Following immunostaining, DAPI staining was performed to highlight the dermal–epidermal junction. Quantification of β-tubulin III + , IB4 + , and CGRP + nerve fibers showed that WT and Prmt6 –/– mice had similar levels of peripheral nerve density. (D) Sciatic nerves from WT and Prmt6 –/– mice were harvested and immunostained with the pan-neural marker β-tubulin III (red) and DAPI (blue). Scale bars: 200 µm in A, B, D and 100 µm in C. Quantification indicated that WT and Prmt6 –/– mice exhibited similar levels of peripheral nerve density. Unpaired t-test was used for all statistical comparisons. Five sections per mouse from four mice per group were evaluated. CGRP: Calcitonin gene-related peptide; DAPI: 4′,6-diamidino-2-phenylindole; DRG: dorsal root ganglion; IB4: isolectin B4; PRMT6: protein arginine methyltransferase-6; Prmt6 –/– : PRMT6 knockout; WT: wild-type.
Article Snippet: The sections were blocked overnight in 5% BSA and 1% Triton X-100 in PBS and then incubated with primary antibodies against PRMT6 (rabbit, 1:200; Novus Biologicals, Littleton, CO, USA, Cat# NB100-56642, RRID: AB_838734), heterogeneous nuclear ribonucleoprotein F (hnRNP-F) (mouse,1:200; Thermo Fisher Scientific, Waltham, MA, USA, Cat# MA5-18024, RRID: AB_2539408), calcitonin gene-related peptide (CGRP; mouse, 1:200, Abcam, Cambridge, MA, USA, Cat# ab81887, RRID: AB_1658411),
Techniques: Marker, Immunostaining, Staining, Knock-Out
Journal: Neural Regeneration Research
Article Title: Protein arginine methyltransferase-6 regulates heterogeneous nuclear ribonucleoprotein-F expression and is a potential target for the treatment of neuropathic pain
doi: 10.4103/NRR.NRR-D-23-01539
Figure Lengend Snippet: hnRNP-F and PRMT6 co-localize in DRG neurons. (A) Immunofluorescence images showing co-localization of hnRNP-F (red) and PRMT6 (green) in DRG neuronal nuclei. (B) Distribution of hnRNP-F (red) within DRG neurons in Prmt6 knockout mice. Approximately 40% of β-tubulin III–positive neurons (green) were also positive for hnRNP-F immunofluorescence in WT mice. Prmt6 knockout in the DRG increased the proportion of cells exhibiting β-tubulin III and hnRNP-F colocalization to 69%. There was no significant change in the relative proportions of different neuron types. Approximately 29% of hnRNP-F-positive neurons were positive for CGRP (green), 40% for IB4 (green), and 27% for NF200 (green) in WT mice, while in Prmt6 knockout mice, approximately 30% of hnRNP-F-positive neurons were positive for CGRP, 40% for IB4, and 30% for NF200. Scale bars: 50 µm in A and B. Unpaired t -test was used. *** P < 0.001, vs . WT. Five sections per mouse from three mice per group were evaluated.
Article Snippet: The sections were blocked overnight in 5% BSA and 1% Triton X-100 in PBS and then incubated with primary antibodies against PRMT6 (rabbit, 1:200; Novus Biologicals, Littleton, CO, USA, Cat# NB100-56642, RRID: AB_838734), heterogeneous nuclear ribonucleoprotein F (hnRNP-F) (mouse,1:200; Thermo Fisher Scientific, Waltham, MA, USA, Cat# MA5-18024, RRID: AB_2539408), calcitonin gene-related peptide (CGRP; mouse, 1:200, Abcam, Cambridge, MA, USA, Cat# ab81887, RRID: AB_1658411),
Techniques: Immunofluorescence, Knock-Out
Journal: Neural Regeneration Research
Article Title: Complement-dependent neuroinflammation in spinal cord injury: from pathology to therapeutic implications
doi: 10.4103/NRR.NRR-D-24-00116
Figure Lengend Snippet: Overview of complement activation pathways. Created with BioRender.com. C: Complement; DAMPs: damage-associated molecular patterns; MBL: mannose-binding lectins; PAMPs: pathogen associated molecular patterns.
Article Snippet: The pattern of activation of the lectin pathway is similar and is initiated when soluble defense proteins like ficolins and mannose-binding
Techniques: Activation Assay, Binding Assay
Journal: Neural Regeneration Research
Article Title: Complement-dependent neuroinflammation in spinal cord injury: from pathology to therapeutic implications
doi: 10.4103/NRR.NRR-D-24-00116
Figure Lengend Snippet: Triggers of complement activation after spinal cord injury. Complement pathways are activated by either the direct binding of opsonins to stressed and degenerating cells, or by antibodies that recognize DAMPs. An alternative mechanism also involves binding to CRP (an acute phase reactant). These mechanisms involve the classical and alternative pathways of C. The lectin pathway has not yet been investigated. Created with BioRender.com. C: Complement; CRP: C-reactive protein; DAMPs: damage-associated molecular patterns; MBL: mannose-binding lectins.
Article Snippet: The pattern of activation of the lectin pathway is similar and is initiated when soluble defense proteins like ficolins and mannose-binding
Techniques: Activation Assay, Binding Assay
Journal: Neural Regeneration Research
Article Title: Complement-dependent neuroinflammation in spinal cord injury: from pathology to therapeutic implications
doi: 10.4103/NRR.NRR-D-24-00116
Figure Lengend Snippet: Inhibitors of complement activation after spinal cord injury. The figure shows the different pathways of C activation and the sites of action of different endogenous and exogenous inhibitors. (1) Inhibitors of the antibody-complement axis include genetic models of Rag1 deficiency, B-cell depletion, and Fc-gamma receptor in addition to the fusion protein B4-Crry that inhibits the bind of B4IgM to modified annexin IV. (2) Inhibitors of the classical pathway include genetic deficiency in C1q or use of C1-inhibitor (C1-INH). (3) Inhibitors of C3 convertase from all pathways include the fusion proteins B4-Crry and CR2-Crry, soluble CR1 (sCR1), and C3 deficiency. (4) Inhibition of the alternative pathway C3 convertase via factor B deficiency. (5) Inhibition of C3a-C3aR1 interaction via C3aR1 deficiency. (6) Inhibition of C5a-C5aR interaction using C5aR1 antagonist (PMX-205), C5aR1 deficiency or C5aR2 deficiency. (7) Inhibition of membrane attack complex includes C5 deficiency and C6 deficiency. Created with BioRender.com. C: Complement; DAMPs: damage-associated molecular patterns; MBL: mannose-binding lectins; PAMPs: pathogen associated molecular patterns.
Article Snippet: The pattern of activation of the lectin pathway is similar and is initiated when soluble defense proteins like ficolins and mannose-binding
Techniques: Activation Assay, Modification, Inhibition, Membrane, Binding Assay
Journal: Neural Regeneration Research
Article Title: Complement-dependent neuroinflammation in spinal cord injury: from pathology to therapeutic implications
doi: 10.4103/NRR.NRR-D-24-00116
Figure Lengend Snippet: Triggers of complement activation after spinal cord injury. Complement pathways are activated by either the direct binding of opsonins to stressed and degenerating cells, or by antibodies that recognize DAMPs. An alternative mechanism also involves binding to CRP (an acute phase reactant). These mechanisms involve the classical and alternative pathways of C. The lectin pathway has not yet been investigated. Created with BioRender.com. C: Complement; CRP: C-reactive protein; DAMPs: damage-associated molecular patterns; MBL: mannose-binding lectins.
Article Snippet: The pattern of activation of the
Techniques: Activation Assay, Binding Assay
Journal: Scientific Reports
Article Title: Drug-resistant epilepsy associated with peripheral complement decreases and sex-specific cytokine imbalances: a pilot study
doi: 10.1038/s41598-025-88654-5
Figure Lengend Snippet: Serum complement component concentrations in patients with drug-resistant epilepsy (DRE) and healthy controls. ( A ) The diagram illustrates the convergence of the three complement pathways: alternative, classical, and lectin, leading to the cleavage of C3 and C5. The alternative pathway is activated through spontaneous hydrolysis of C3. The formation of C3b and Bb components (from Factor B) allows the cleavage of C3, being regulated by Factor H. All the pathways converge in the cleavage of C5 into C5b which helps assemble the membrane-attack complex (MAC) (terminal pathway). ( B ) Volcano plot illustrates the differential expression of complement proteins between DRE patients and healthy controls. The y-axis represents -log10(p-values) from t-tests and the x-axis represents log2 fold change (FC). ( C-O ) Serum concentrations of multiple complement molecules in DRE patients (blue) and healthy controls (gray): C1q, Mannose-binding Lectin (MBL), Factor B, Factor H, C2, C4, C4b, C3, C3a, C3b/iC3b, C5, C5a, and C5b-9. Violin plots show individual data points, median, and quartiles. Statistical analysis was performed using an unpaired t -test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
Article Snippet: In healthy individuals (Fig. A,C), we observed high positive correlations between classical (C1q to C3b/iC3b; females, r = 0.72, p < 0.001; males, r = 0.89, p < 0.001),
Techniques: Membrane, Expressing, Binding Assay
Journal: Scientific Reports
Article Title: Drug-resistant epilepsy associated with peripheral complement decreases and sex-specific cytokine imbalances: a pilot study
doi: 10.1038/s41598-025-88654-5
Figure Lengend Snippet: Serum complement component concentrations in male and female patients with drug-resistant epilepsy (DRE) and healthy controls. ( A-C ) Volcano plots illustrate the differential expression of complement proteins between male or female healthy and DRE groups. The y-axis represents -log10(p-values) from two-way ANOVA, and the x-axis represents log2 fold change (FC). ( A ) Healthy females and healthy males ( B ) DRE females and healthy females ( C ) DRE males and healthy males. ( D-P ) Serum concentrations of multiple complement molecules: C1q, Mannose-binding Lectin (MBL), Factor B, Factor H, C2, C4, C4b, C3, C3a, C3b/iC3b, C5, C5a, and C5b-9. DRE patients are represented in blue, and healthy controls in gray. Violin plots display individual data points, median, and quartiles. Statistical analysis was performed using two-way ANOVA with categorical variables: condition (DRE or healthy control) and sex (female and male). Significance levels: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
Article Snippet: In healthy individuals (Fig. A,C), we observed high positive correlations between classical (C1q to C3b/iC3b; females, r = 0.72, p < 0.001; males, r = 0.89, p < 0.001),
Techniques: Expressing, Binding Assay, Control
Journal: Scientific Reports
Article Title: Drug-resistant epilepsy associated with peripheral complement decreases and sex-specific cytokine imbalances: a pilot study
doi: 10.1038/s41598-025-88654-5
Figure Lengend Snippet: Correlations between complement components. ( A-D ) Pearson correlation matrices of all complement components in ( A ) Healthy females, ( B ) DRE females, ( C ) Healthy males, and ( D ) DRE males. ( E-H ) Pearson correlations between C1q and C3b/iC3b in ( E ) Healthy females, ( F ) DRE females, ( G ) Healthy males, and ( H ) DRE males. I-L , Pearson correlations between MBL and C3b/iC3b in ( I ) Healthy females, ( J ) DRE females, ( K ) Healthy males, and ( L ) DRE males. ( M-P ) Pearson correlations between C5a and C3b/iC3b in ( M ) Healthy females, ( N ) DRE females, ( O ) Healthy males, and ( P ) DRE males. Gray areas in panels ( E-P ) indicate 95% confidence intervals. Top: Simplified diagram showing the classical (C1q) and lectin (MBL) pathways and downstream cleavage of C3 and C5. Red indicates correlations shown in panels ( E-P ). Supplementary Figs. 1–4 show all r and p values for all correlations by group.
Article Snippet: In healthy individuals (Fig. A,C), we observed high positive correlations between classical (C1q to C3b/iC3b; females, r = 0.72, p < 0.001; males, r = 0.89, p < 0.001),
Techniques:
Journal: Scientific Reports
Article Title: Drug-resistant epilepsy associated with peripheral complement decreases and sex-specific cytokine imbalances: a pilot study
doi: 10.1038/s41598-025-88654-5
Figure Lengend Snippet: Correlations between serum concentrations of complement components and cytokines. A-D , Pearson correlation matrices between complement components and cytokines in ( A ) Healthy females, ( B ) DRE females, ( C ) Healthy males, and ( D ) DRE males. E-H , Pearson correlations between IL-17 and C3b/iC3b in ( E ) Healthy females, ( F ) DRE females, ( G ) Healthy males, and ( H ) DRE males. I-L , Pearson correlations between IL-8 and C4 in ( I ) Healthy females, ( J ) DRE females, ( K ) Healthy males, and ( L ) DRE males. M-P , Pearson correlations between CCL2 and C3b/iC3b in ( M ) Healthy females, ( N ) DRE females, ( O ) Healthy males, and ( P ) DRE males. Gray areas in panels E-P indicate 95% confidence intervals. Supplementary Figs. 1–4 show all r and p values for all correlations by group.
Article Snippet: In healthy individuals (Fig. A,C), we observed high positive correlations between classical (C1q to C3b/iC3b; females, r = 0.72, p < 0.001; males, r = 0.89, p < 0.001),
Techniques: