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    Alomone Labs kv1 3
    Fyn modulates the posttranslational modification of <t>Kv1.3.</t> (A) Western blot analysis of postmortem human PD and age-matched control brains showing increased phosphorylation of Kv1.3. (B) Immunoprecipitation of Fyn and Kv1.3 showing direct Fyn-Kv1.3 interaction after αSyn Agg treatment. ( C ) Duolink PLA showing αSyn Agg -induced interaction between Kv1.3 and Fyn. Scale bar: 25 μm. ( D ) Western blot of Fyn WT and KO PMCs revealed that Kv1.3 phosphorylation at residue 135 was Fyn dependent. ( E ) IHC analysis of substantia nigra from Fyn +/+ and Fyn –/– mice showing reduced phosphorylation of Kv1.3 after αSyn PFF injection. Scale bars: 100 μm; 60 μm (insets). ( F ) IHC of substantia nigra from MitoPark mice and their littermate controls showing that pharmacological inhibition of Fyn by saracatinib reduced Kv1.3 phosphorylation. Scale bar: 100 μm. ( G – J ) Immortalized MMCs were either transfected with WT Kv1.3 or aY135A Kv1.3 plasmid. ( G ) qRT-PCR analysis and ( H ) Griess assay showing reduced levels of inducible NOS (iNOS) and nitrite release, respectively, in Y135A Kv1.3-transfected cells compared with WT cells. ( I ) qRT-PCR analysis showing reduced IL-1β production in Y135A Kv1.3–transfected versus WT Kv1.3–transfected MMCs. ( J ) Luminex assay showing reduced IL-1β secretion in Y135A Kv1.3–transfected compared with WT Kv1.3–transfected MMCs. A 1-way ANOVA was used to compare multiple groups. In most cases, Tukey’s post hoc analysis was applied. A 2-tailed Student’s t test was used to compare 2 groups in A . Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–4 biological replicates from 2–3 independent experiments. * P ≤ 0.05, ** P
    Kv1 3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    NeuroMab kv1 3
    Callosal <t>Kv1.3</t> channel protein in axons and glia is altered after injury and with CFZ treatment A. Confocal overlay showing Kv1.3 ( red ) and Kv1.2 ( green ) in rat corpus callosum 24h following midline fluid percussion TBI. Low magnification shows that each channel protein is found in reactive glia around callosal vessels (arrowheads) and along axon bundles (arrows). Inset shows paired paranodal distribution of Kv1.3 and Kv1.2 channels, some nodes with co-localization (yellow arrow), others with single channel expression (green, red arrows). B . Confocal overlays showing Kv1.3 in callosal astrocytes of sham injured (GFAP+, left-arrows; inset shows cell body and perivascular co-localization) and microglia of 24h postinjury cases (IBA1+, right-arrows). C. Western blot (WB) of protein extracts from 24h postinjury corpus callosum revealed that TBI reduced 67kD Kv1.3 levels and that CFZ treatment reversed loss of Kv1.3 expression. Data expressed as percent of paired untreated sham controls run on same blot. Lanes representative of group effects are shown in each panel. (ANOVA, *p
    Kv1 3, supplied by NeuroMab, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/kv1 3/product/NeuroMab
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    93
    Alomone Labs anti kv1 3
    Callosal <t>Kv1.3</t> channel protein in axons and glia is altered after injury and with CFZ treatment A. Confocal overlay showing Kv1.3 ( red ) and Kv1.2 ( green ) in rat corpus callosum 24h following midline fluid percussion TBI. Low magnification shows that each channel protein is found in reactive glia around callosal vessels (arrowheads) and along axon bundles (arrows). Inset shows paired paranodal distribution of Kv1.3 and Kv1.2 channels, some nodes with co-localization (yellow arrow), others with single channel expression (green, red arrows). B . Confocal overlays showing Kv1.3 in callosal astrocytes of sham injured (GFAP+, left-arrows; inset shows cell body and perivascular co-localization) and microglia of 24h postinjury cases (IBA1+, right-arrows). C. Western blot (WB) of protein extracts from 24h postinjury corpus callosum revealed that TBI reduced 67kD Kv1.3 levels and that CFZ treatment reversed loss of Kv1.3 expression. Data expressed as percent of paired untreated sham controls run on same blot. Lanes representative of group effects are shown in each panel. (ANOVA, *p
    Anti Kv1 3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Jackson Immuno kv1 3
    K + -channel expression in acutely isolated microglia. (a) <t>Kv1.3</t> current density increases in microglia from the infarct area after MCAO (28.8 ± 2.0 pA/pF, n = 19) and microglia isolated from the hippocampus following intracerebroventricular LPS injection (22.9 ± 16.6 pA/pF, n = 13) compared to microglia from wild-type control brains (5.0 ± 3.9 pA/pF, n = 16) or microglia from the contralateral side after MCAO (5.7 ± 4.4 pA/pF, n = 18). (b) Example current traces showing Kv1.3's characteristic use-dependence and sensitivity to the Kv1.3 blockers PAP-1 and ShK-L5. (c) Microglia from the contralateral (50.2 ± 35.4 pS/pF, n = 18) and ipsilateral side after MCAO (71.6 ± 34.9 pS/pF, n = 21), as well as microglia isolated from the hippocampus following intracerebroventricular LPS injection (84.0 ± 42.4 pS/pF, n = 13) show higher KCa3.1 current densities than microglia from wild-type control brains (29.7 ± 15.2 pS/pF, n = 16). (d) Example KCa3.1 current traces elicited by a ramp protocol showing the current's sensitive to 1 µM of the KCa3.1-selective blocker TRAM-34. (e) Microglia from both the contralateral side (7.8 ± 5.8 pA/pF, n = 18) and the infarct area (15.1 ± 10.2 pA/pF, n = 21) after MCAO show increased Kir current densities compared to those from wild-type (2.4 ± 2.4 pA/pF, n = 16) or LPS-injected brains (1.9 ± 2.9 pA/pF, n = 13). (f) Representative current traces showing a large Kir current, which was observable in some MCAO microglia, but not in microglia isolated from the hippocampus following intracerebroventricular LPS injection. Data are presented as mean ± S.D. Statistical significance was determined by Student's t -test.
    Kv1 3, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Fyn modulates the posttranslational modification of Kv1.3. (A) Western blot analysis of postmortem human PD and age-matched control brains showing increased phosphorylation of Kv1.3. (B) Immunoprecipitation of Fyn and Kv1.3 showing direct Fyn-Kv1.3 interaction after αSyn Agg treatment. ( C ) Duolink PLA showing αSyn Agg -induced interaction between Kv1.3 and Fyn. Scale bar: 25 μm. ( D ) Western blot of Fyn WT and KO PMCs revealed that Kv1.3 phosphorylation at residue 135 was Fyn dependent. ( E ) IHC analysis of substantia nigra from Fyn +/+ and Fyn –/– mice showing reduced phosphorylation of Kv1.3 after αSyn PFF injection. Scale bars: 100 μm; 60 μm (insets). ( F ) IHC of substantia nigra from MitoPark mice and their littermate controls showing that pharmacological inhibition of Fyn by saracatinib reduced Kv1.3 phosphorylation. Scale bar: 100 μm. ( G – J ) Immortalized MMCs were either transfected with WT Kv1.3 or aY135A Kv1.3 plasmid. ( G ) qRT-PCR analysis and ( H ) Griess assay showing reduced levels of inducible NOS (iNOS) and nitrite release, respectively, in Y135A Kv1.3-transfected cells compared with WT cells. ( I ) qRT-PCR analysis showing reduced IL-1β production in Y135A Kv1.3–transfected versus WT Kv1.3–transfected MMCs. ( J ) Luminex assay showing reduced IL-1β secretion in Y135A Kv1.3–transfected compared with WT Kv1.3–transfected MMCs. A 1-way ANOVA was used to compare multiple groups. In most cases, Tukey’s post hoc analysis was applied. A 2-tailed Student’s t test was used to compare 2 groups in A . Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–4 biological replicates from 2–3 independent experiments. * P ≤ 0.05, ** P

    Journal: The Journal of Clinical Investigation

    Article Title: Kv1.3 modulates neuroinflammation and neurodegeneration in Parkinson’s disease

    doi: 10.1172/JCI136174

    Figure Lengend Snippet: Fyn modulates the posttranslational modification of Kv1.3. (A) Western blot analysis of postmortem human PD and age-matched control brains showing increased phosphorylation of Kv1.3. (B) Immunoprecipitation of Fyn and Kv1.3 showing direct Fyn-Kv1.3 interaction after αSyn Agg treatment. ( C ) Duolink PLA showing αSyn Agg -induced interaction between Kv1.3 and Fyn. Scale bar: 25 μm. ( D ) Western blot of Fyn WT and KO PMCs revealed that Kv1.3 phosphorylation at residue 135 was Fyn dependent. ( E ) IHC analysis of substantia nigra from Fyn +/+ and Fyn –/– mice showing reduced phosphorylation of Kv1.3 after αSyn PFF injection. Scale bars: 100 μm; 60 μm (insets). ( F ) IHC of substantia nigra from MitoPark mice and their littermate controls showing that pharmacological inhibition of Fyn by saracatinib reduced Kv1.3 phosphorylation. Scale bar: 100 μm. ( G – J ) Immortalized MMCs were either transfected with WT Kv1.3 or aY135A Kv1.3 plasmid. ( G ) qRT-PCR analysis and ( H ) Griess assay showing reduced levels of inducible NOS (iNOS) and nitrite release, respectively, in Y135A Kv1.3-transfected cells compared with WT cells. ( I ) qRT-PCR analysis showing reduced IL-1β production in Y135A Kv1.3–transfected versus WT Kv1.3–transfected MMCs. ( J ) Luminex assay showing reduced IL-1β secretion in Y135A Kv1.3–transfected compared with WT Kv1.3–transfected MMCs. A 1-way ANOVA was used to compare multiple groups. In most cases, Tukey’s post hoc analysis was applied. A 2-tailed Student’s t test was used to compare 2 groups in A . Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–4 biological replicates from 2–3 independent experiments. * P ≤ 0.05, ** P

    Article Snippet: Kv1.3 in psoriatic disease: PAP-1, a small molecule inhibitor of Kv1.3 is effective in the SCID mouse psoriasis--xenograft model. J Autoimmun.

    Techniques: Modification, Western Blot, Immunoprecipitation, Proximity Ligation Assay, Immunohistochemistry, Mouse Assay, Injection, Inhibition, Transfection, Plasmid Preparation, Quantitative RT-PCR, Griess Assay, Luminex

    Kv1.3 inhibition protects against αSyn PFF -induced behavior deficit and dopaminergic neuronal loss. ( A ) Treatment paradigm corresponding to the αSyn PFF mouse model of PD. ( B ) Representative movement tracks showing that PAP-1 rescued movement deficits induced by αSyn PFF . ( C – E ) A VersaMax open-field test showed decreased ( C ) rest time and increased ( D ) horizontal activity and ( E ) total distance traveled for αSyn PFF mice treated with PAP-1. ( F and G ) HPLC showing that PAP-1 treatment protected against loss of ( F ) dopamine and ( G ) DOPAC induced by αSyn PFF . ( H ) Western blot analysis of TH showing loss of TH induced by αSyn PFF in the SNpc region. A 1-way ANOVA was used to compare multiple groups. In most cases, Tukey’s post hoc analysis was applied. Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–7 animals per group. * P ≤ 0.05, ** P

    Journal: The Journal of Clinical Investigation

    Article Title: Kv1.3 modulates neuroinflammation and neurodegeneration in Parkinson’s disease

    doi: 10.1172/JCI136174

    Figure Lengend Snippet: Kv1.3 inhibition protects against αSyn PFF -induced behavior deficit and dopaminergic neuronal loss. ( A ) Treatment paradigm corresponding to the αSyn PFF mouse model of PD. ( B ) Representative movement tracks showing that PAP-1 rescued movement deficits induced by αSyn PFF . ( C – E ) A VersaMax open-field test showed decreased ( C ) rest time and increased ( D ) horizontal activity and ( E ) total distance traveled for αSyn PFF mice treated with PAP-1. ( F and G ) HPLC showing that PAP-1 treatment protected against loss of ( F ) dopamine and ( G ) DOPAC induced by αSyn PFF . ( H ) Western blot analysis of TH showing loss of TH induced by αSyn PFF in the SNpc region. A 1-way ANOVA was used to compare multiple groups. In most cases, Tukey’s post hoc analysis was applied. Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–7 animals per group. * P ≤ 0.05, ** P

    Article Snippet: Kv1.3 in psoriatic disease: PAP-1, a small molecule inhibitor of Kv1.3 is effective in the SCID mouse psoriasis--xenograft model. J Autoimmun.

    Techniques: Inhibition, Activity Assay, Mouse Assay, High Performance Liquid Chromatography, Western Blot

    Upregulated expression of the potassium channel Kv1.3 upon aggregated αSyn stimulation in ex vivo slices and B cells derived from patients with PD. ( A ) Midbrain slice cultures were treated with 1 μM αSyn Agg for 24 hours. qRT-PCR shows upregulated Kv1.3 mRNA expression. ( B ) Western blot shows upregulated Kv1.3 protein level in midbrain slice cultures treated with 1 μM αSyn Agg for 24 hours. ( C ) qRT-PCR of midbrain slice cultures treated with 1 μM αSyn Agg for 24 hours, revealing upregulation of the proinflammatory factors Nos2 , Csf2 , IL-6 , IL-1β , and Tnfa . ( D ) qRT-PCR shows increased Kv1.3 mRNA expression in B cell lymphocytes isolated from patients with PD compared with expression in B cell lymphocytes from age-matched controls. ( E ) Whole-cell patch clamping of B cell lymphocytes isolated from patients with PD showed higher Kv1.3 channel activity compared with that observed in age-matched controls ( n = 3 control and n = 3 PD). A 1-way ANOVA was used to compare multiple groups in C and D . Tukey’s post hoc analysis was applied. A 2-tailed Student’s t test was used to compare 2 groups. Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–7 biological replicates from 2–3 independent experiments unless otherwise indicated. * P ≤ 0.05 and ** P

    Journal: The Journal of Clinical Investigation

    Article Title: Kv1.3 modulates neuroinflammation and neurodegeneration in Parkinson’s disease

    doi: 10.1172/JCI136174

    Figure Lengend Snippet: Upregulated expression of the potassium channel Kv1.3 upon aggregated αSyn stimulation in ex vivo slices and B cells derived from patients with PD. ( A ) Midbrain slice cultures were treated with 1 μM αSyn Agg for 24 hours. qRT-PCR shows upregulated Kv1.3 mRNA expression. ( B ) Western blot shows upregulated Kv1.3 protein level in midbrain slice cultures treated with 1 μM αSyn Agg for 24 hours. ( C ) qRT-PCR of midbrain slice cultures treated with 1 μM αSyn Agg for 24 hours, revealing upregulation of the proinflammatory factors Nos2 , Csf2 , IL-6 , IL-1β , and Tnfa . ( D ) qRT-PCR shows increased Kv1.3 mRNA expression in B cell lymphocytes isolated from patients with PD compared with expression in B cell lymphocytes from age-matched controls. ( E ) Whole-cell patch clamping of B cell lymphocytes isolated from patients with PD showed higher Kv1.3 channel activity compared with that observed in age-matched controls ( n = 3 control and n = 3 PD). A 1-way ANOVA was used to compare multiple groups in C and D . Tukey’s post hoc analysis was applied. A 2-tailed Student’s t test was used to compare 2 groups. Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–7 biological replicates from 2–3 independent experiments unless otherwise indicated. * P ≤ 0.05 and ** P

    Article Snippet: Kv1.3 in psoriatic disease: PAP-1, a small molecule inhibitor of Kv1.3 is effective in the SCID mouse psoriasis--xenograft model. J Autoimmun.

    Techniques: Expressing, Ex Vivo, Derivative Assay, Quantitative RT-PCR, Western Blot, Isolation, Activity Assay

    Kv1.3 expression is highly induced in microglial cells in experimental models of PD and postmortem PD brains. ( A ) Western blot showing increased Kv1.3 protein levels in the substantia nigra of the Syn-AAV mouse model of PD. ( B ) qRT-PCR analysis of 8- to 24-week-old nigral tissues from the MitoPark mouse model of PD showing Kv1.3 induction compared with age-matched littermate controls. ( C ) Western blot of 24-week-old nigral tissues from the MitoPark mouse model of PD (MP) showing induction of Kv1.3 protein expression compared with age-matched littermate control mice (LM). ( D ) IHC in 24-week-old nigral tissues from the MitoPark mouse model of PD showing higher Kv1.3 protein levels (red) in IBA1-positive microglial cells (green) compared with age-matched controls as revealed by their colocalization (yellow). Scale bar: 20 μm. ( E ) qRT-PCR analysis of nigral tissues from the MPTP mouse model revealing induction of Kv1.3 mRNA expression. ( F ) Western blot showing increased Kv1.3 protein levels in substantia nigra of the MPTP mouse model of PD. ( G ) qRT-PCR analysis of postmortem human PD brains showing elevated Kv1.3 mRNA expression. ( H ) Western blot of the SN region of postmortem human PD brain showing induction of Kv1.3 protein expression compared with age-matched controls. n = 6–8. ( I ) Immunostaining revealing higher Kv1.3 levels in the prefrontal cortex of postmortem human PD brains compared with age-matched controls. Lower panel shows the deconvoluted binary image used for analysis. Three regions per brain were analyzed. Scale bar: 200 μm. ( J ) Dual DAB staining showing induction of Kv1.3 expression in HLA-DR–positive microglial cells in patients with DLBs compared with age-matched controls. Scale bars: 100 μm; 20 μm (enlarged insets). A 1-way ANOVA was used to compare multiple groups. Tukey’s post hoc analysis was applied B . A 2-tailed Student’s t test was used to compare 2 groups. Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–9 biological replicates from 2–3 independent experiments unless otherwise indicated. * P ≤ 0.05, ** P

    Journal: The Journal of Clinical Investigation

    Article Title: Kv1.3 modulates neuroinflammation and neurodegeneration in Parkinson’s disease

    doi: 10.1172/JCI136174

    Figure Lengend Snippet: Kv1.3 expression is highly induced in microglial cells in experimental models of PD and postmortem PD brains. ( A ) Western blot showing increased Kv1.3 protein levels in the substantia nigra of the Syn-AAV mouse model of PD. ( B ) qRT-PCR analysis of 8- to 24-week-old nigral tissues from the MitoPark mouse model of PD showing Kv1.3 induction compared with age-matched littermate controls. ( C ) Western blot of 24-week-old nigral tissues from the MitoPark mouse model of PD (MP) showing induction of Kv1.3 protein expression compared with age-matched littermate control mice (LM). ( D ) IHC in 24-week-old nigral tissues from the MitoPark mouse model of PD showing higher Kv1.3 protein levels (red) in IBA1-positive microglial cells (green) compared with age-matched controls as revealed by their colocalization (yellow). Scale bar: 20 μm. ( E ) qRT-PCR analysis of nigral tissues from the MPTP mouse model revealing induction of Kv1.3 mRNA expression. ( F ) Western blot showing increased Kv1.3 protein levels in substantia nigra of the MPTP mouse model of PD. ( G ) qRT-PCR analysis of postmortem human PD brains showing elevated Kv1.3 mRNA expression. ( H ) Western blot of the SN region of postmortem human PD brain showing induction of Kv1.3 protein expression compared with age-matched controls. n = 6–8. ( I ) Immunostaining revealing higher Kv1.3 levels in the prefrontal cortex of postmortem human PD brains compared with age-matched controls. Lower panel shows the deconvoluted binary image used for analysis. Three regions per brain were analyzed. Scale bar: 200 μm. ( J ) Dual DAB staining showing induction of Kv1.3 expression in HLA-DR–positive microglial cells in patients with DLBs compared with age-matched controls. Scale bars: 100 μm; 20 μm (enlarged insets). A 1-way ANOVA was used to compare multiple groups. Tukey’s post hoc analysis was applied B . A 2-tailed Student’s t test was used to compare 2 groups. Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–9 biological replicates from 2–3 independent experiments unless otherwise indicated. * P ≤ 0.05, ** P

    Article Snippet: Kv1.3 in psoriatic disease: PAP-1, a small molecule inhibitor of Kv1.3 is effective in the SCID mouse psoriasis--xenograft model. J Autoimmun.

    Techniques: Expressing, Western Blot, Quantitative RT-PCR, Mouse Assay, Immunohistochemistry, Immunostaining, Staining

    Fyn modulates the transcriptional regulation of Kv1.3 in microglial cells through the Fyn/PKCδ kinase signaling cascade. ( A ) In silico analysis of the promoter sequence of Kv1.3 revealed probable Nf-κB– and SP1-binding sites. ( B ) qRT-PCR analysis of immortalized MMCs cotreated with αSyn Agg and either SN50 (100 μg/mL) or SB203580 (1 μM), showing that both compounds attenuated αSyn Agg -induced Kv1.3 expression. ( C ) Western blot of Fyn WT and KO PMCs treated with αSyn Agg , showing that Fyn KO reduced the induction of the p38 MAPK pathway. ( D ) qRT-PCR analysis revealed that Fyn KO reduced αSyn Agg -induced Kv1.3 mRNA levels. ( E ) Whole-cell patch-clamp recording showing that Fyn KO attenuated αSyn Agg - and LPS-induced Kv1.3 activity compared with Fyn WT PMCs (WT control n = 24, WT αSyn Agg n = 12, WT LPS n = 29, Fyn KO αSyn Agg n = 20, Fyn KO LPS n = 15). ( F ) ICC showing that Fyn KO reduced αSyn Agg -induced Kv1.3 protein levels in PMCs. Scale bar: 15 μm. ( G ) ICC of PMCs revealed that αSyn Agg -induced Kv1.3 protein expression was reduced by PKCδ KO. Scale bar: 15 μm. ( H ) qRT-PCR analysis of PMCs showing that PKC KO reduced the expression of αSyn Agg -induced Kv1.3 mRNA. ( I ) Whole-cell patch clam recording of PMCs showing that PKC KO attenuated αSyn Agg - and LPS-induced Kv1.3 activity compared with PKC WT PMCs (WT control n = 24, WT αSyn Agg n = 12, WT LPS n = 20, PKC-KO αSyn Agg n = 29, PKC-KO LPS n = 35). Data are presented as the mean ± SD. A 1-way ANOVA was used to compare multiple groups. In most cases, Tukey’s post hoc analysis was applied. Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–4 biological replicates from 2–3 independent experiments unless otherwise indicated. * P ≤ 0.05, ** P

    Journal: The Journal of Clinical Investigation

    Article Title: Kv1.3 modulates neuroinflammation and neurodegeneration in Parkinson’s disease

    doi: 10.1172/JCI136174

    Figure Lengend Snippet: Fyn modulates the transcriptional regulation of Kv1.3 in microglial cells through the Fyn/PKCδ kinase signaling cascade. ( A ) In silico analysis of the promoter sequence of Kv1.3 revealed probable Nf-κB– and SP1-binding sites. ( B ) qRT-PCR analysis of immortalized MMCs cotreated with αSyn Agg and either SN50 (100 μg/mL) or SB203580 (1 μM), showing that both compounds attenuated αSyn Agg -induced Kv1.3 expression. ( C ) Western blot of Fyn WT and KO PMCs treated with αSyn Agg , showing that Fyn KO reduced the induction of the p38 MAPK pathway. ( D ) qRT-PCR analysis revealed that Fyn KO reduced αSyn Agg -induced Kv1.3 mRNA levels. ( E ) Whole-cell patch-clamp recording showing that Fyn KO attenuated αSyn Agg - and LPS-induced Kv1.3 activity compared with Fyn WT PMCs (WT control n = 24, WT αSyn Agg n = 12, WT LPS n = 29, Fyn KO αSyn Agg n = 20, Fyn KO LPS n = 15). ( F ) ICC showing that Fyn KO reduced αSyn Agg -induced Kv1.3 protein levels in PMCs. Scale bar: 15 μm. ( G ) ICC of PMCs revealed that αSyn Agg -induced Kv1.3 protein expression was reduced by PKCδ KO. Scale bar: 15 μm. ( H ) qRT-PCR analysis of PMCs showing that PKC KO reduced the expression of αSyn Agg -induced Kv1.3 mRNA. ( I ) Whole-cell patch clam recording of PMCs showing that PKC KO attenuated αSyn Agg - and LPS-induced Kv1.3 activity compared with PKC WT PMCs (WT control n = 24, WT αSyn Agg n = 12, WT LPS n = 20, PKC-KO αSyn Agg n = 29, PKC-KO LPS n = 35). Data are presented as the mean ± SD. A 1-way ANOVA was used to compare multiple groups. In most cases, Tukey’s post hoc analysis was applied. Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–4 biological replicates from 2–3 independent experiments unless otherwise indicated. * P ≤ 0.05, ** P

    Article Snippet: Kv1.3 in psoriatic disease: PAP-1, a small molecule inhibitor of Kv1.3 is effective in the SCID mouse psoriasis--xenograft model. J Autoimmun.

    Techniques: In Silico, Sequencing, Binding Assay, Quantitative RT-PCR, Expressing, Western Blot, Patch Clamp, Activity Assay, Immunocytochemistry

    Kv1.3 modulates neuroinflammation in a cell culture model of PD. ( A – C ) Kv1.3 WT and KO PMCs were treated with 1 μM αSyn Agg for 24 hours. Luminex analysis shows that Kv1.3 KO reduced the release of the αSyn Agg -induced proinflammatory factors ( A ) TNF-α, ( B ) IL-12, and ( C ) IL-1β. ( D – H ) Immortalized MMCs were transfected with WT a Kv1.3 plasmid, and then 48 hours after transfection, cells were treated with 1 μM αSyn Agg for 24 hours. ( D – F ) qRT-PCR analysis showing that Kv1.3 overexpression aggravated αSyn Agg -induced production of the proinflammatory factors ( D ) Nos2 , ( E ) pro– IL-1β , and ( F ) TNF-α . ( G and H ) Luminex analysis showing that Kv1.3 overexpression potentiated the release of the proinflammatory factors ( G ) IL-6 and ( H ) IL-12. ( I ) Voltage ramp from –120 mV to 40 mV elicited a characteristic outward rectifying current in αSyn Agg -treated microglia that was sensitive to the Kv1.3-selective inhibitor PAP-1. ( J ) LDH assay showing that PAP-1 reduced αSyn Agg -induced LDH release from microglial cells. ( K – M ) Luminex assay revealing that PAP-1 attenuated the αSyn Agg -induced proinflammatory factors ( K ) IL-12, ( L ) TNF-α, and ( M ) IL-6. ( N ) Western blot analysis demonstrating that PAP-1 reduced αSyn Agg -induced NLRP3 expression. ( O ) ICC analysis revealed that PAP-1 reduced NLRP3 expression induced by αSyn Agg . Scale bar: 25 μm. A 1-way ANOVA was performed to compare multiple groups. In most cases, Tukey’s post hoc analysis was applied. Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–4 biological replicates from 2–3 independent experiments. * P ≤ 0.05, ** P

    Journal: The Journal of Clinical Investigation

    Article Title: Kv1.3 modulates neuroinflammation and neurodegeneration in Parkinson’s disease

    doi: 10.1172/JCI136174

    Figure Lengend Snippet: Kv1.3 modulates neuroinflammation in a cell culture model of PD. ( A – C ) Kv1.3 WT and KO PMCs were treated with 1 μM αSyn Agg for 24 hours. Luminex analysis shows that Kv1.3 KO reduced the release of the αSyn Agg -induced proinflammatory factors ( A ) TNF-α, ( B ) IL-12, and ( C ) IL-1β. ( D – H ) Immortalized MMCs were transfected with WT a Kv1.3 plasmid, and then 48 hours after transfection, cells were treated with 1 μM αSyn Agg for 24 hours. ( D – F ) qRT-PCR analysis showing that Kv1.3 overexpression aggravated αSyn Agg -induced production of the proinflammatory factors ( D ) Nos2 , ( E ) pro– IL-1β , and ( F ) TNF-α . ( G and H ) Luminex analysis showing that Kv1.3 overexpression potentiated the release of the proinflammatory factors ( G ) IL-6 and ( H ) IL-12. ( I ) Voltage ramp from –120 mV to 40 mV elicited a characteristic outward rectifying current in αSyn Agg -treated microglia that was sensitive to the Kv1.3-selective inhibitor PAP-1. ( J ) LDH assay showing that PAP-1 reduced αSyn Agg -induced LDH release from microglial cells. ( K – M ) Luminex assay revealing that PAP-1 attenuated the αSyn Agg -induced proinflammatory factors ( K ) IL-12, ( L ) TNF-α, and ( M ) IL-6. ( N ) Western blot analysis demonstrating that PAP-1 reduced αSyn Agg -induced NLRP3 expression. ( O ) ICC analysis revealed that PAP-1 reduced NLRP3 expression induced by αSyn Agg . Scale bar: 25 μm. A 1-way ANOVA was performed to compare multiple groups. In most cases, Tukey’s post hoc analysis was applied. Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–4 biological replicates from 2–3 independent experiments. * P ≤ 0.05, ** P

    Article Snippet: Kv1.3 in psoriatic disease: PAP-1, a small molecule inhibitor of Kv1.3 is effective in the SCID mouse psoriasis--xenograft model. J Autoimmun.

    Techniques: Cell Culture, Luminex, Transfection, Plasmid Preparation, Quantitative RT-PCR, Over Expression, Lactate Dehydrogenase Assay, Western Blot, Expressing, Immunocytochemistry

    Upregulated expression of the potassium channel Kv1.3 upon aggregated αSyn stimulation in microglial cells in vitro. ( A ) Whole-cell patch-clamp recordings of PMCs treated with 1 μM αSyn Agg for 24–48 hours, showing αSyn Agg -induced increased Kv1.3 activity (control n = 24 and αSyn Agg n = 12). Kv1.3 was identified by its characteristic use dependence, which was revealed when applying a train of ten 200-ms pulses from –80 to 40 mV at 1-second intervals (1 Hz). ( B ) qRT-PCR showing that αSyn Agg induced Kv1.3 mRNA expression without significantly altering other potassium channels. ( C ) Western blot of αSyn Agg -induced Kv1.3 protein expression in PMCs. ( D ) ICC of αSyn Agg -induced Kv1.3 protein expression in PMCs. Scale bar: 100 μm. ( E ) Flow cytometric analysis of immortalized MMCs treated with 1 μM αSyn Agg for 24 hours, showing αSyn Agg -induced Kv1.3 surface expression. ( F ) qRT-PCR of human microglia treated with LPS (1 μg/mL) and IL-4 (20 ng/mL) for 6 hours, showing LPS-induced Kv1.3 expression. A 1-way ANOVA was used to compare multiple groups. In most cases, Tukey’s post hoc analysis was applied in B and F . A 2-tailed Student’s t test was used for all other figures when comparing 2 groups. Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–5 biological replicates from 2–3 independent experiments unless otherwise noted. * P ≤ 0.05, ** P

    Journal: The Journal of Clinical Investigation

    Article Title: Kv1.3 modulates neuroinflammation and neurodegeneration in Parkinson’s disease

    doi: 10.1172/JCI136174

    Figure Lengend Snippet: Upregulated expression of the potassium channel Kv1.3 upon aggregated αSyn stimulation in microglial cells in vitro. ( A ) Whole-cell patch-clamp recordings of PMCs treated with 1 μM αSyn Agg for 24–48 hours, showing αSyn Agg -induced increased Kv1.3 activity (control n = 24 and αSyn Agg n = 12). Kv1.3 was identified by its characteristic use dependence, which was revealed when applying a train of ten 200-ms pulses from –80 to 40 mV at 1-second intervals (1 Hz). ( B ) qRT-PCR showing that αSyn Agg induced Kv1.3 mRNA expression without significantly altering other potassium channels. ( C ) Western blot of αSyn Agg -induced Kv1.3 protein expression in PMCs. ( D ) ICC of αSyn Agg -induced Kv1.3 protein expression in PMCs. Scale bar: 100 μm. ( E ) Flow cytometric analysis of immortalized MMCs treated with 1 μM αSyn Agg for 24 hours, showing αSyn Agg -induced Kv1.3 surface expression. ( F ) qRT-PCR of human microglia treated with LPS (1 μg/mL) and IL-4 (20 ng/mL) for 6 hours, showing LPS-induced Kv1.3 expression. A 1-way ANOVA was used to compare multiple groups. In most cases, Tukey’s post hoc analysis was applied in B and F . A 2-tailed Student’s t test was used for all other figures when comparing 2 groups. Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–5 biological replicates from 2–3 independent experiments unless otherwise noted. * P ≤ 0.05, ** P

    Article Snippet: Kv1.3 in psoriatic disease: PAP-1, a small molecule inhibitor of Kv1.3 is effective in the SCID mouse psoriasis--xenograft model. J Autoimmun.

    Techniques: Expressing, In Vitro, Patch Clamp, Activity Assay, Quantitative RT-PCR, Western Blot, Immunocytochemistry

    Callosal Kv1.3 channel protein in axons and glia is altered after injury and with CFZ treatment A. Confocal overlay showing Kv1.3 ( red ) and Kv1.2 ( green ) in rat corpus callosum 24h following midline fluid percussion TBI. Low magnification shows that each channel protein is found in reactive glia around callosal vessels (arrowheads) and along axon bundles (arrows). Inset shows paired paranodal distribution of Kv1.3 and Kv1.2 channels, some nodes with co-localization (yellow arrow), others with single channel expression (green, red arrows). B . Confocal overlays showing Kv1.3 in callosal astrocytes of sham injured (GFAP+, left-arrows; inset shows cell body and perivascular co-localization) and microglia of 24h postinjury cases (IBA1+, right-arrows). C. Western blot (WB) of protein extracts from 24h postinjury corpus callosum revealed that TBI reduced 67kD Kv1.3 levels and that CFZ treatment reversed loss of Kv1.3 expression. Data expressed as percent of paired untreated sham controls run on same blot. Lanes representative of group effects are shown in each panel. (ANOVA, *p

    Journal: Experimental neurology

    Article Title: Targeting Kv1.3 channels to reduce white matter pathology after traumatic brain injury

    doi: 10.1016/j.expneurol.2016.06.011

    Figure Lengend Snippet: Callosal Kv1.3 channel protein in axons and glia is altered after injury and with CFZ treatment A. Confocal overlay showing Kv1.3 ( red ) and Kv1.2 ( green ) in rat corpus callosum 24h following midline fluid percussion TBI. Low magnification shows that each channel protein is found in reactive glia around callosal vessels (arrowheads) and along axon bundles (arrows). Inset shows paired paranodal distribution of Kv1.3 and Kv1.2 channels, some nodes with co-localization (yellow arrow), others with single channel expression (green, red arrows). B . Confocal overlays showing Kv1.3 in callosal astrocytes of sham injured (GFAP+, left-arrows; inset shows cell body and perivascular co-localization) and microglia of 24h postinjury cases (IBA1+, right-arrows). C. Western blot (WB) of protein extracts from 24h postinjury corpus callosum revealed that TBI reduced 67kD Kv1.3 levels and that CFZ treatment reversed loss of Kv1.3 expression. Data expressed as percent of paired untreated sham controls run on same blot. Lanes representative of group effects are shown in each panel. (ANOVA, *p

    Article Snippet: This pattern of results, a CFZ treatment effect on Kv1.3, but not Kv1.2 protein, is consistent with the prior finding that CFZ is highly selective for Kv1.3, relative to Kv1.2 channels.

    Techniques: Expressing, Western Blot

    Corpus callosum mixed glial cultures grown in MatTek dishes and subjected to confocal dual labeling with antibodies to Kv1.3 (green) and microglial marker protein IBA1 (red) or astrocyte marker protein GFAP (red) A. Microglia are predominantly ramified in untreated cultures (arrowhead), shifting to reactive rounded cells with lobular processes after LPS (arrows). Kv1.3 protein is localized within the majority of microglia in each field and shifts from a normal concentration around nuclei (inset, UNT), to a more uniform distribution after inflammatory stimulation (inset, LPS). B. Astrocytes show mixed flat and fibrous morphologies. Kv1.3 signal is much reduced in astrocytes relative to surrounding microglia (not stained in these images) and predominantly found in small aggregates around cell nuclei (arrow). Astrocytes do not show LPS group differences in Kv1.3 expression. Bars= 20 µm.

    Journal: Experimental neurology

    Article Title: Targeting Kv1.3 channels to reduce white matter pathology after traumatic brain injury

    doi: 10.1016/j.expneurol.2016.06.011

    Figure Lengend Snippet: Corpus callosum mixed glial cultures grown in MatTek dishes and subjected to confocal dual labeling with antibodies to Kv1.3 (green) and microglial marker protein IBA1 (red) or astrocyte marker protein GFAP (red) A. Microglia are predominantly ramified in untreated cultures (arrowhead), shifting to reactive rounded cells with lobular processes after LPS (arrows). Kv1.3 protein is localized within the majority of microglia in each field and shifts from a normal concentration around nuclei (inset, UNT), to a more uniform distribution after inflammatory stimulation (inset, LPS). B. Astrocytes show mixed flat and fibrous morphologies. Kv1.3 signal is much reduced in astrocytes relative to surrounding microglia (not stained in these images) and predominantly found in small aggregates around cell nuclei (arrow). Astrocytes do not show LPS group differences in Kv1.3 expression. Bars= 20 µm.

    Article Snippet: This pattern of results, a CFZ treatment effect on Kv1.3, but not Kv1.2 protein, is consistent with the prior finding that CFZ is highly selective for Kv1.3, relative to Kv1.2 channels.

    Techniques: Labeling, Marker, Concentration Assay, Staining, Expressing

    K + -channel expression in acutely isolated microglia. (a) Kv1.3 current density increases in microglia from the infarct area after MCAO (28.8 ± 2.0 pA/pF, n = 19) and microglia isolated from the hippocampus following intracerebroventricular LPS injection (22.9 ± 16.6 pA/pF, n = 13) compared to microglia from wild-type control brains (5.0 ± 3.9 pA/pF, n = 16) or microglia from the contralateral side after MCAO (5.7 ± 4.4 pA/pF, n = 18). (b) Example current traces showing Kv1.3's characteristic use-dependence and sensitivity to the Kv1.3 blockers PAP-1 and ShK-L5. (c) Microglia from the contralateral (50.2 ± 35.4 pS/pF, n = 18) and ipsilateral side after MCAO (71.6 ± 34.9 pS/pF, n = 21), as well as microglia isolated from the hippocampus following intracerebroventricular LPS injection (84.0 ± 42.4 pS/pF, n = 13) show higher KCa3.1 current densities than microglia from wild-type control brains (29.7 ± 15.2 pS/pF, n = 16). (d) Example KCa3.1 current traces elicited by a ramp protocol showing the current's sensitive to 1 µM of the KCa3.1-selective blocker TRAM-34. (e) Microglia from both the contralateral side (7.8 ± 5.8 pA/pF, n = 18) and the infarct area (15.1 ± 10.2 pA/pF, n = 21) after MCAO show increased Kir current densities compared to those from wild-type (2.4 ± 2.4 pA/pF, n = 16) or LPS-injected brains (1.9 ± 2.9 pA/pF, n = 13). (f) Representative current traces showing a large Kir current, which was observable in some MCAO microglia, but not in microglia isolated from the hippocampus following intracerebroventricular LPS injection. Data are presented as mean ± S.D. Statistical significance was determined by Student's t -test.

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: The potassium channel KCa3.1 constitutes a pharmacological target for neuroinflammation associated with ischemia/reperfusion stroke

    doi: 10.1177/0271678X15611434

    Figure Lengend Snippet: K + -channel expression in acutely isolated microglia. (a) Kv1.3 current density increases in microglia from the infarct area after MCAO (28.8 ± 2.0 pA/pF, n = 19) and microglia isolated from the hippocampus following intracerebroventricular LPS injection (22.9 ± 16.6 pA/pF, n = 13) compared to microglia from wild-type control brains (5.0 ± 3.9 pA/pF, n = 16) or microglia from the contralateral side after MCAO (5.7 ± 4.4 pA/pF, n = 18). (b) Example current traces showing Kv1.3's characteristic use-dependence and sensitivity to the Kv1.3 blockers PAP-1 and ShK-L5. (c) Microglia from the contralateral (50.2 ± 35.4 pS/pF, n = 18) and ipsilateral side after MCAO (71.6 ± 34.9 pS/pF, n = 21), as well as microglia isolated from the hippocampus following intracerebroventricular LPS injection (84.0 ± 42.4 pS/pF, n = 13) show higher KCa3.1 current densities than microglia from wild-type control brains (29.7 ± 15.2 pS/pF, n = 16). (d) Example KCa3.1 current traces elicited by a ramp protocol showing the current's sensitive to 1 µM of the KCa3.1-selective blocker TRAM-34. (e) Microglia from both the contralateral side (7.8 ± 5.8 pA/pF, n = 18) and the infarct area (15.1 ± 10.2 pA/pF, n = 21) after MCAO show increased Kir current densities compared to those from wild-type (2.4 ± 2.4 pA/pF, n = 16) or LPS-injected brains (1.9 ± 2.9 pA/pF, n = 13). (f) Representative current traces showing a large Kir current, which was observable in some MCAO microglia, but not in microglia isolated from the hippocampus following intracerebroventricular LPS injection. Data are presented as mean ± S.D. Statistical significance was determined by Student's t -test.

    Article Snippet: In contrast, microglia isolated from the infarcted area after MCAO or from the hippocampus of adult mice 48 h after intracerebroventricular LPS injection were clearly activated based on their increased cell size (measured as capacity) and showed roughly five-fold larger Kv currents ( a), which seemed to be predominantly carried by Kv1.3 based on their use-dependence and sensitivity to the specific Kv1.3 blockers PAP-1 and ShK-L5 ( b).

    Techniques: Expressing, Isolation, Injection

    KCa3.1 and Kv1.3 are expressed on microglia/macrophages in human infarcts. (a) KCa3.1 staining in a 2–3-week-old infarct. KCa3.1 expression is localized to macrophages/microglia (M) and vascular endothelial (E) cells. (b) Fluorescent staining for a microglia/macrophage marker (MAC387) and KCa3.1. (c) Kv1.3 staining in a 14-day old-infarct. (d) Fluorescent staining for a microglia/macrophage marker (MAC387) and Kv1.3. All images are from 5-μm thick paraffin sections.

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: The potassium channel KCa3.1 constitutes a pharmacological target for neuroinflammation associated with ischemia/reperfusion stroke

    doi: 10.1177/0271678X15611434

    Figure Lengend Snippet: KCa3.1 and Kv1.3 are expressed on microglia/macrophages in human infarcts. (a) KCa3.1 staining in a 2–3-week-old infarct. KCa3.1 expression is localized to macrophages/microglia (M) and vascular endothelial (E) cells. (b) Fluorescent staining for a microglia/macrophage marker (MAC387) and KCa3.1. (c) Kv1.3 staining in a 14-day old-infarct. (d) Fluorescent staining for a microglia/macrophage marker (MAC387) and Kv1.3. All images are from 5-μm thick paraffin sections.

    Article Snippet: In contrast, microglia isolated from the infarcted area after MCAO or from the hippocampus of adult mice 48 h after intracerebroventricular LPS injection were clearly activated based on their increased cell size (measured as capacity) and showed roughly five-fold larger Kv currents ( a), which seemed to be predominantly carried by Kv1.3 based on their use-dependence and sensitivity to the specific Kv1.3 blockers PAP-1 and ShK-L5 ( b).

    Techniques: Staining, Expressing, Marker