hcn2  (Alomone Labs)


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  • 86

    Structured Review

    Alomone Labs hcn2
    Primary antibodies.
    Hcn2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hcn2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hcn2 - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons"

    Article Title: High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons

    Journal: Cells

    doi: 10.3390/cells13110944

    Primary antibodies.
    Figure Legend Snippet: Primary antibodies.

    Techniques Used: Concentration Assay

    Characterization of the molecular interactions between Cav1.3, SK3, and HCN channels using co-immunoprecipitation and immunoblot. Proteins were membrane-extracted in CL47a solubilization buffer (IP1) or in CL48 buffer (IP2) and immunoprecipitated using anti-Cav1.3 ( A ), anti-SK3 ( B ), and anti-Kv4.3 antibodies ( C ). Blots were performed on starting material, P2 membrane fraction (P2), or on solubilized P2 proteins (sol) or after immunoprecipitation (IP1, IP2). Blots were probed with anti-SK3 (( A ), right panel), anti-Cav1.3 (( B ), right panel), anti-HCN4 (( A , B ) left panels; ( C ), right panel), anti-Kv4.3 (( C ), left panel), and anti-HCN2 (( C ), middle panel). P2: enriched P2 membrane fraction. IP1: immunoprecipitated proteins in CL47a buffer. IP2: immunoprecipitated proteins in CL48 buffer. sol: solubilisate. WB: Western blot (immunoblot).
    Figure Legend Snippet: Characterization of the molecular interactions between Cav1.3, SK3, and HCN channels using co-immunoprecipitation and immunoblot. Proteins were membrane-extracted in CL47a solubilization buffer (IP1) or in CL48 buffer (IP2) and immunoprecipitated using anti-Cav1.3 ( A ), anti-SK3 ( B ), and anti-Kv4.3 antibodies ( C ). Blots were performed on starting material, P2 membrane fraction (P2), or on solubilized P2 proteins (sol) or after immunoprecipitation (IP1, IP2). Blots were probed with anti-SK3 (( A ), right panel), anti-Cav1.3 (( B ), right panel), anti-HCN4 (( A , B ) left panels; ( C ), right panel), anti-Kv4.3 (( C ), left panel), and anti-HCN2 (( C ), middle panel). P2: enriched P2 membrane fraction. IP1: immunoprecipitated proteins in CL47a buffer. IP2: immunoprecipitated proteins in CL48 buffer. sol: solubilisate. WB: Western blot (immunoblot).

    Techniques Used: Immunoprecipitation, Western Blot, Membrane

    Protein nano-environment of midbrain ion channels determined using affinity-purification mass spectrometry. ( A ) Left panel, Cav1.2 subunit ( Cacna1c gene) immunoprecipitations. Right panel, Cav1.3 subunit ( Cacna1d gene) immunoprecipitation. ( B ) SK3 subunit ( Kcnn3 gene) immunoprecipitation. ( C ) Left panel, HCN2 subunit immunoprecipitation. Right panel, HCN4 subunit immunoprecipitation. ( D ) Kv4.3 ( Kcnd3 gene) immunoprecipitation. High-confidence Class A (solid line) and low-confidence Class B (dashed lines) thresholds are displayed on the plots. Plain squares represent the bait and empty square interactants. The most relevant of the significant interactants are named.
    Figure Legend Snippet: Protein nano-environment of midbrain ion channels determined using affinity-purification mass spectrometry. ( A ) Left panel, Cav1.2 subunit ( Cacna1c gene) immunoprecipitations. Right panel, Cav1.3 subunit ( Cacna1d gene) immunoprecipitation. ( B ) SK3 subunit ( Kcnn3 gene) immunoprecipitation. ( C ) Left panel, HCN2 subunit immunoprecipitation. Right panel, HCN4 subunit immunoprecipitation. ( D ) Kv4.3 ( Kcnd3 gene) immunoprecipitation. High-confidence Class A (solid line) and low-confidence Class B (dashed lines) thresholds are displayed on the plots. Plain squares represent the bait and empty square interactants. The most relevant of the significant interactants are named.

    Techniques Used: Affinity Purification, Mass Spectrometry, Immunoprecipitation

    Visualization of the mass-spectrometry-based protein interaction network. The 6 ion channels of interest are depicted using different colors (blue, Kv4.3; pink and red, HCN2 and HCN4; yellow and orange, Cav1.2 and Cav1.3; green, SK3). Interactions are shown with lines of according colors. Thick lines represent Class A interactions, thinner lines Class B. Putative interactants were compared with the CRAPome 2.0 database (Contaminant Repository for Affinity Purification). Proteins identified in less than 1% of all CRAPome negative experiments were considered highly specific (black circles), between 1 and 2%, specific (grey circles), and between 2 and 10%, weakly specific (white circles) interactants.
    Figure Legend Snippet: Visualization of the mass-spectrometry-based protein interaction network. The 6 ion channels of interest are depicted using different colors (blue, Kv4.3; pink and red, HCN2 and HCN4; yellow and orange, Cav1.2 and Cav1.3; green, SK3). Interactions are shown with lines of according colors. Thick lines represent Class A interactions, thinner lines Class B. Putative interactants were compared with the CRAPome 2.0 database (Contaminant Repository for Affinity Purification). Proteins identified in less than 1% of all CRAPome negative experiments were considered highly specific (black circles), between 1 and 2%, specific (grey circles), and between 2 and 10%, weakly specific (white circles) interactants.

    Techniques Used: Mass Spectrometry, Affinity Purification

    HCN2, HCN4, and Sarm1 expression during midbrain postnatal development. Immunoblots of membrane-enriched fraction from adult (Ad) and 9 (P9) or 11 (P11) postnatal days Wt and Sarm1 −/− mice. Blots were probed with anti-HCN2 (left panel), anti-HCN4 (middle panel), or anti-Sarm1 (right panel) antibodies. WB: Western blot (immunoblot).
    Figure Legend Snippet: HCN2, HCN4, and Sarm1 expression during midbrain postnatal development. Immunoblots of membrane-enriched fraction from adult (Ad) and 9 (P9) or 11 (P11) postnatal days Wt and Sarm1 −/− mice. Blots were probed with anti-HCN2 (left panel), anti-HCN4 (middle panel), or anti-Sarm1 (right panel) antibodies. WB: Western blot (immunoblot).

    Techniques Used: Expressing, Western Blot, Membrane

    Cav1.3-HCN2-SK3 complex characterization in SNc DA neurons using proximity ligation assay. ( A ) Left, representative images of the PLA reaction (green) in wild-type (Wt) or Cav1.3 −/− SNc DA neurons using anti-Cav1.3 and anti-HCN2 antibodies. Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( B ) Representative images of the PLA reaction (green) in wild-type (Wt) SNc DA neurons using anti-SK3 and anti-HCN2 antibodies (top row) or anti-SK2 and anti-HCN2 antibodies (bottom row). Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( C ) Quantification of PLA fluorescent puncta on SNc DA neurons. Left, Cav1.3 and HCN2 PLA from 3 Wt (empty black circles; n = 21 images) or 3 Cav1.3 −/− (empty red circles; n = 18 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Middle, HCN2 and SK3 PLA (empty black circles; n = 11 images) or HCN2 and SK2 (empty orange circles; n = 7 images) from 2 independent experiments or anti-Kv4.3 and HCN2 (empty grey circle; n = 4 images) PLA from 1 experiment. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Right, Cav1.3 and SK3 PLA from 2 Wt (empty black circles; n = 10 images) or 2 Cav1.3 −/− (empty red circles; n = 8 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. ** p < 0.005; *** p < 0.001, Wilcoxon–Mann–Whitney test.
    Figure Legend Snippet: Cav1.3-HCN2-SK3 complex characterization in SNc DA neurons using proximity ligation assay. ( A ) Left, representative images of the PLA reaction (green) in wild-type (Wt) or Cav1.3 −/− SNc DA neurons using anti-Cav1.3 and anti-HCN2 antibodies. Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( B ) Representative images of the PLA reaction (green) in wild-type (Wt) SNc DA neurons using anti-SK3 and anti-HCN2 antibodies (top row) or anti-SK2 and anti-HCN2 antibodies (bottom row). Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( C ) Quantification of PLA fluorescent puncta on SNc DA neurons. Left, Cav1.3 and HCN2 PLA from 3 Wt (empty black circles; n = 21 images) or 3 Cav1.3 −/− (empty red circles; n = 18 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Middle, HCN2 and SK3 PLA (empty black circles; n = 11 images) or HCN2 and SK2 (empty orange circles; n = 7 images) from 2 independent experiments or anti-Kv4.3 and HCN2 (empty grey circle; n = 4 images) PLA from 1 experiment. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Right, Cav1.3 and SK3 PLA from 2 Wt (empty black circles; n = 10 images) or 2 Cav1.3 −/− (empty red circles; n = 8 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. ** p < 0.005; *** p < 0.001, Wilcoxon–Mann–Whitney test.

    Techniques Used: Proximity Ligation Assay, Immunolabeling, MANN-WHITNEY

    hcn2  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
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  • 86

    Structured Review

    Alomone Labs hcn2
    Primary antibodies.
    Hcn2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hcn2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hcn2 - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons"

    Article Title: High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons

    Journal: Cells

    doi: 10.3390/cells13110944

    Primary antibodies.
    Figure Legend Snippet: Primary antibodies.

    Techniques Used: Concentration Assay

    Characterization of the molecular interactions between Cav1.3, SK3, and HCN channels using co-immunoprecipitation and immunoblot. Proteins were membrane-extracted in CL47a solubilization buffer (IP1) or in CL48 buffer (IP2) and immunoprecipitated using anti-Cav1.3 ( A ), anti-SK3 ( B ), and anti-Kv4.3 antibodies ( C ). Blots were performed on starting material, P2 membrane fraction (P2), or on solubilized P2 proteins (sol) or after immunoprecipitation (IP1, IP2). Blots were probed with anti-SK3 (( A ), right panel), anti-Cav1.3 (( B ), right panel), anti-HCN4 (( A , B ) left panels; ( C ), right panel), anti-Kv4.3 (( C ), left panel), and anti-HCN2 (( C ), middle panel). P2: enriched P2 membrane fraction. IP1: immunoprecipitated proteins in CL47a buffer. IP2: immunoprecipitated proteins in CL48 buffer. sol: solubilisate. WB: Western blot (immunoblot).
    Figure Legend Snippet: Characterization of the molecular interactions between Cav1.3, SK3, and HCN channels using co-immunoprecipitation and immunoblot. Proteins were membrane-extracted in CL47a solubilization buffer (IP1) or in CL48 buffer (IP2) and immunoprecipitated using anti-Cav1.3 ( A ), anti-SK3 ( B ), and anti-Kv4.3 antibodies ( C ). Blots were performed on starting material, P2 membrane fraction (P2), or on solubilized P2 proteins (sol) or after immunoprecipitation (IP1, IP2). Blots were probed with anti-SK3 (( A ), right panel), anti-Cav1.3 (( B ), right panel), anti-HCN4 (( A , B ) left panels; ( C ), right panel), anti-Kv4.3 (( C ), left panel), and anti-HCN2 (( C ), middle panel). P2: enriched P2 membrane fraction. IP1: immunoprecipitated proteins in CL47a buffer. IP2: immunoprecipitated proteins in CL48 buffer. sol: solubilisate. WB: Western blot (immunoblot).

    Techniques Used: Immunoprecipitation, Western Blot, Membrane

    Protein nano-environment of midbrain ion channels determined using affinity-purification mass spectrometry. ( A ) Left panel, Cav1.2 subunit ( Cacna1c gene) immunoprecipitations. Right panel, Cav1.3 subunit ( Cacna1d gene) immunoprecipitation. ( B ) SK3 subunit ( Kcnn3 gene) immunoprecipitation. ( C ) Left panel, HCN2 subunit immunoprecipitation. Right panel, HCN4 subunit immunoprecipitation. ( D ) Kv4.3 ( Kcnd3 gene) immunoprecipitation. High-confidence Class A (solid line) and low-confidence Class B (dashed lines) thresholds are displayed on the plots. Plain squares represent the bait and empty square interactants. The most relevant of the significant interactants are named.
    Figure Legend Snippet: Protein nano-environment of midbrain ion channels determined using affinity-purification mass spectrometry. ( A ) Left panel, Cav1.2 subunit ( Cacna1c gene) immunoprecipitations. Right panel, Cav1.3 subunit ( Cacna1d gene) immunoprecipitation. ( B ) SK3 subunit ( Kcnn3 gene) immunoprecipitation. ( C ) Left panel, HCN2 subunit immunoprecipitation. Right panel, HCN4 subunit immunoprecipitation. ( D ) Kv4.3 ( Kcnd3 gene) immunoprecipitation. High-confidence Class A (solid line) and low-confidence Class B (dashed lines) thresholds are displayed on the plots. Plain squares represent the bait and empty square interactants. The most relevant of the significant interactants are named.

    Techniques Used: Affinity Purification, Mass Spectrometry, Immunoprecipitation

    Visualization of the mass-spectrometry-based protein interaction network. The 6 ion channels of interest are depicted using different colors (blue, Kv4.3; pink and red, HCN2 and HCN4; yellow and orange, Cav1.2 and Cav1.3; green, SK3). Interactions are shown with lines of according colors. Thick lines represent Class A interactions, thinner lines Class B. Putative interactants were compared with the CRAPome 2.0 database (Contaminant Repository for Affinity Purification). Proteins identified in less than 1% of all CRAPome negative experiments were considered highly specific (black circles), between 1 and 2%, specific (grey circles), and between 2 and 10%, weakly specific (white circles) interactants.
    Figure Legend Snippet: Visualization of the mass-spectrometry-based protein interaction network. The 6 ion channels of interest are depicted using different colors (blue, Kv4.3; pink and red, HCN2 and HCN4; yellow and orange, Cav1.2 and Cav1.3; green, SK3). Interactions are shown with lines of according colors. Thick lines represent Class A interactions, thinner lines Class B. Putative interactants were compared with the CRAPome 2.0 database (Contaminant Repository for Affinity Purification). Proteins identified in less than 1% of all CRAPome negative experiments were considered highly specific (black circles), between 1 and 2%, specific (grey circles), and between 2 and 10%, weakly specific (white circles) interactants.

    Techniques Used: Mass Spectrometry, Affinity Purification

    HCN2, HCN4, and Sarm1 expression during midbrain postnatal development. Immunoblots of membrane-enriched fraction from adult (Ad) and 9 (P9) or 11 (P11) postnatal days Wt and Sarm1 −/− mice. Blots were probed with anti-HCN2 (left panel), anti-HCN4 (middle panel), or anti-Sarm1 (right panel) antibodies. WB: Western blot (immunoblot).
    Figure Legend Snippet: HCN2, HCN4, and Sarm1 expression during midbrain postnatal development. Immunoblots of membrane-enriched fraction from adult (Ad) and 9 (P9) or 11 (P11) postnatal days Wt and Sarm1 −/− mice. Blots were probed with anti-HCN2 (left panel), anti-HCN4 (middle panel), or anti-Sarm1 (right panel) antibodies. WB: Western blot (immunoblot).

    Techniques Used: Expressing, Western Blot, Membrane

    Cav1.3-HCN2-SK3 complex characterization in SNc DA neurons using proximity ligation assay. ( A ) Left, representative images of the PLA reaction (green) in wild-type (Wt) or Cav1.3 −/− SNc DA neurons using anti-Cav1.3 and anti-HCN2 antibodies. Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( B ) Representative images of the PLA reaction (green) in wild-type (Wt) SNc DA neurons using anti-SK3 and anti-HCN2 antibodies (top row) or anti-SK2 and anti-HCN2 antibodies (bottom row). Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( C ) Quantification of PLA fluorescent puncta on SNc DA neurons. Left, Cav1.3 and HCN2 PLA from 3 Wt (empty black circles; n = 21 images) or 3 Cav1.3 −/− (empty red circles; n = 18 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Middle, HCN2 and SK3 PLA (empty black circles; n = 11 images) or HCN2 and SK2 (empty orange circles; n = 7 images) from 2 independent experiments or anti-Kv4.3 and HCN2 (empty grey circle; n = 4 images) PLA from 1 experiment. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Right, Cav1.3 and SK3 PLA from 2 Wt (empty black circles; n = 10 images) or 2 Cav1.3 −/− (empty red circles; n = 8 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. ** p < 0.005; *** p < 0.001, Wilcoxon–Mann–Whitney test.
    Figure Legend Snippet: Cav1.3-HCN2-SK3 complex characterization in SNc DA neurons using proximity ligation assay. ( A ) Left, representative images of the PLA reaction (green) in wild-type (Wt) or Cav1.3 −/− SNc DA neurons using anti-Cav1.3 and anti-HCN2 antibodies. Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( B ) Representative images of the PLA reaction (green) in wild-type (Wt) SNc DA neurons using anti-SK3 and anti-HCN2 antibodies (top row) or anti-SK2 and anti-HCN2 antibodies (bottom row). Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( C ) Quantification of PLA fluorescent puncta on SNc DA neurons. Left, Cav1.3 and HCN2 PLA from 3 Wt (empty black circles; n = 21 images) or 3 Cav1.3 −/− (empty red circles; n = 18 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Middle, HCN2 and SK3 PLA (empty black circles; n = 11 images) or HCN2 and SK2 (empty orange circles; n = 7 images) from 2 independent experiments or anti-Kv4.3 and HCN2 (empty grey circle; n = 4 images) PLA from 1 experiment. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Right, Cav1.3 and SK3 PLA from 2 Wt (empty black circles; n = 10 images) or 2 Cav1.3 −/− (empty red circles; n = 8 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. ** p < 0.005; *** p < 0.001, Wilcoxon–Mann–Whitney test.

    Techniques Used: Proximity Ligation Assay, Immunolabeling, MANN-WHITNEY

    hcn2  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Alomone Labs hcn2
    Primary antibodies.
    Hcn2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hcn2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hcn2 - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons"

    Article Title: High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons

    Journal: Cells

    doi: 10.3390/cells13110944

    Primary antibodies.
    Figure Legend Snippet: Primary antibodies.

    Techniques Used: Concentration Assay

    Characterization of the molecular interactions between Cav1.3, SK3, and HCN channels using co-immunoprecipitation and immunoblot. Proteins were membrane-extracted in CL47a solubilization buffer (IP1) or in CL48 buffer (IP2) and immunoprecipitated using anti-Cav1.3 ( A ), anti-SK3 ( B ), and anti-Kv4.3 antibodies ( C ). Blots were performed on starting material, P2 membrane fraction (P2), or on solubilized P2 proteins (sol) or after immunoprecipitation (IP1, IP2). Blots were probed with anti-SK3 (( A ), right panel), anti-Cav1.3 (( B ), right panel), anti-HCN4 (( A , B ) left panels; ( C ), right panel), anti-Kv4.3 (( C ), left panel), and anti-HCN2 (( C ), middle panel). P2: enriched P2 membrane fraction. IP1: immunoprecipitated proteins in CL47a buffer. IP2: immunoprecipitated proteins in CL48 buffer. sol: solubilisate. WB: Western blot (immunoblot).
    Figure Legend Snippet: Characterization of the molecular interactions between Cav1.3, SK3, and HCN channels using co-immunoprecipitation and immunoblot. Proteins were membrane-extracted in CL47a solubilization buffer (IP1) or in CL48 buffer (IP2) and immunoprecipitated using anti-Cav1.3 ( A ), anti-SK3 ( B ), and anti-Kv4.3 antibodies ( C ). Blots were performed on starting material, P2 membrane fraction (P2), or on solubilized P2 proteins (sol) or after immunoprecipitation (IP1, IP2). Blots were probed with anti-SK3 (( A ), right panel), anti-Cav1.3 (( B ), right panel), anti-HCN4 (( A , B ) left panels; ( C ), right panel), anti-Kv4.3 (( C ), left panel), and anti-HCN2 (( C ), middle panel). P2: enriched P2 membrane fraction. IP1: immunoprecipitated proteins in CL47a buffer. IP2: immunoprecipitated proteins in CL48 buffer. sol: solubilisate. WB: Western blot (immunoblot).

    Techniques Used: Immunoprecipitation, Western Blot, Membrane

    Protein nano-environment of midbrain ion channels determined using affinity-purification mass spectrometry. ( A ) Left panel, Cav1.2 subunit ( Cacna1c gene) immunoprecipitations. Right panel, Cav1.3 subunit ( Cacna1d gene) immunoprecipitation. ( B ) SK3 subunit ( Kcnn3 gene) immunoprecipitation. ( C ) Left panel, HCN2 subunit immunoprecipitation. Right panel, HCN4 subunit immunoprecipitation. ( D ) Kv4.3 ( Kcnd3 gene) immunoprecipitation. High-confidence Class A (solid line) and low-confidence Class B (dashed lines) thresholds are displayed on the plots. Plain squares represent the bait and empty square interactants. The most relevant of the significant interactants are named.
    Figure Legend Snippet: Protein nano-environment of midbrain ion channels determined using affinity-purification mass spectrometry. ( A ) Left panel, Cav1.2 subunit ( Cacna1c gene) immunoprecipitations. Right panel, Cav1.3 subunit ( Cacna1d gene) immunoprecipitation. ( B ) SK3 subunit ( Kcnn3 gene) immunoprecipitation. ( C ) Left panel, HCN2 subunit immunoprecipitation. Right panel, HCN4 subunit immunoprecipitation. ( D ) Kv4.3 ( Kcnd3 gene) immunoprecipitation. High-confidence Class A (solid line) and low-confidence Class B (dashed lines) thresholds are displayed on the plots. Plain squares represent the bait and empty square interactants. The most relevant of the significant interactants are named.

    Techniques Used: Affinity Purification, Mass Spectrometry, Immunoprecipitation

    Visualization of the mass-spectrometry-based protein interaction network. The 6 ion channels of interest are depicted using different colors (blue, Kv4.3; pink and red, HCN2 and HCN4; yellow and orange, Cav1.2 and Cav1.3; green, SK3). Interactions are shown with lines of according colors. Thick lines represent Class A interactions, thinner lines Class B. Putative interactants were compared with the CRAPome 2.0 database (Contaminant Repository for Affinity Purification). Proteins identified in less than 1% of all CRAPome negative experiments were considered highly specific (black circles), between 1 and 2%, specific (grey circles), and between 2 and 10%, weakly specific (white circles) interactants.
    Figure Legend Snippet: Visualization of the mass-spectrometry-based protein interaction network. The 6 ion channels of interest are depicted using different colors (blue, Kv4.3; pink and red, HCN2 and HCN4; yellow and orange, Cav1.2 and Cav1.3; green, SK3). Interactions are shown with lines of according colors. Thick lines represent Class A interactions, thinner lines Class B. Putative interactants were compared with the CRAPome 2.0 database (Contaminant Repository for Affinity Purification). Proteins identified in less than 1% of all CRAPome negative experiments were considered highly specific (black circles), between 1 and 2%, specific (grey circles), and between 2 and 10%, weakly specific (white circles) interactants.

    Techniques Used: Mass Spectrometry, Affinity Purification

    HCN2, HCN4, and Sarm1 expression during midbrain postnatal development. Immunoblots of membrane-enriched fraction from adult (Ad) and 9 (P9) or 11 (P11) postnatal days Wt and Sarm1 −/− mice. Blots were probed with anti-HCN2 (left panel), anti-HCN4 (middle panel), or anti-Sarm1 (right panel) antibodies. WB: Western blot (immunoblot).
    Figure Legend Snippet: HCN2, HCN4, and Sarm1 expression during midbrain postnatal development. Immunoblots of membrane-enriched fraction from adult (Ad) and 9 (P9) or 11 (P11) postnatal days Wt and Sarm1 −/− mice. Blots were probed with anti-HCN2 (left panel), anti-HCN4 (middle panel), or anti-Sarm1 (right panel) antibodies. WB: Western blot (immunoblot).

    Techniques Used: Expressing, Western Blot, Membrane

    Cav1.3-HCN2-SK3 complex characterization in SNc DA neurons using proximity ligation assay. ( A ) Left, representative images of the PLA reaction (green) in wild-type (Wt) or Cav1.3 −/− SNc DA neurons using anti-Cav1.3 and anti-HCN2 antibodies. Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( B ) Representative images of the PLA reaction (green) in wild-type (Wt) SNc DA neurons using anti-SK3 and anti-HCN2 antibodies (top row) or anti-SK2 and anti-HCN2 antibodies (bottom row). Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( C ) Quantification of PLA fluorescent puncta on SNc DA neurons. Left, Cav1.3 and HCN2 PLA from 3 Wt (empty black circles; n = 21 images) or 3 Cav1.3 −/− (empty red circles; n = 18 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Middle, HCN2 and SK3 PLA (empty black circles; n = 11 images) or HCN2 and SK2 (empty orange circles; n = 7 images) from 2 independent experiments or anti-Kv4.3 and HCN2 (empty grey circle; n = 4 images) PLA from 1 experiment. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Right, Cav1.3 and SK3 PLA from 2 Wt (empty black circles; n = 10 images) or 2 Cav1.3 −/− (empty red circles; n = 8 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. ** p < 0.005; *** p < 0.001, Wilcoxon–Mann–Whitney test.
    Figure Legend Snippet: Cav1.3-HCN2-SK3 complex characterization in SNc DA neurons using proximity ligation assay. ( A ) Left, representative images of the PLA reaction (green) in wild-type (Wt) or Cav1.3 −/− SNc DA neurons using anti-Cav1.3 and anti-HCN2 antibodies. Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( B ) Representative images of the PLA reaction (green) in wild-type (Wt) SNc DA neurons using anti-SK3 and anti-HCN2 antibodies (top row) or anti-SK2 and anti-HCN2 antibodies (bottom row). Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( C ) Quantification of PLA fluorescent puncta on SNc DA neurons. Left, Cav1.3 and HCN2 PLA from 3 Wt (empty black circles; n = 21 images) or 3 Cav1.3 −/− (empty red circles; n = 18 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Middle, HCN2 and SK3 PLA (empty black circles; n = 11 images) or HCN2 and SK2 (empty orange circles; n = 7 images) from 2 independent experiments or anti-Kv4.3 and HCN2 (empty grey circle; n = 4 images) PLA from 1 experiment. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Right, Cav1.3 and SK3 PLA from 2 Wt (empty black circles; n = 10 images) or 2 Cav1.3 −/− (empty red circles; n = 8 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. ** p < 0.005; *** p < 0.001, Wilcoxon–Mann–Whitney test.

    Techniques Used: Proximity Ligation Assay, Immunolabeling, MANN-WHITNEY

    hcn2  (Alomone Labs)


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    Alomone Labs hcn2
    Primary antibodies.
    Hcn2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hcn2 - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons"

    Article Title: High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons

    Journal: Cells

    doi: 10.3390/cells13110944

    Primary antibodies.
    Figure Legend Snippet: Primary antibodies.

    Techniques Used: Concentration Assay

    Characterization of the molecular interactions between Cav1.3, SK3, and HCN channels using co-immunoprecipitation and immunoblot. Proteins were membrane-extracted in CL47a solubilization buffer (IP1) or in CL48 buffer (IP2) and immunoprecipitated using anti-Cav1.3 ( A ), anti-SK3 ( B ), and anti-Kv4.3 antibodies ( C ). Blots were performed on starting material, P2 membrane fraction (P2), or on solubilized P2 proteins (sol) or after immunoprecipitation (IP1, IP2). Blots were probed with anti-SK3 (( A ), right panel), anti-Cav1.3 (( B ), right panel), anti-HCN4 (( A , B ) left panels; ( C ), right panel), anti-Kv4.3 (( C ), left panel), and anti-HCN2 (( C ), middle panel). P2: enriched P2 membrane fraction. IP1: immunoprecipitated proteins in CL47a buffer. IP2: immunoprecipitated proteins in CL48 buffer. sol: solubilisate. WB: Western blot (immunoblot).
    Figure Legend Snippet: Characterization of the molecular interactions between Cav1.3, SK3, and HCN channels using co-immunoprecipitation and immunoblot. Proteins were membrane-extracted in CL47a solubilization buffer (IP1) or in CL48 buffer (IP2) and immunoprecipitated using anti-Cav1.3 ( A ), anti-SK3 ( B ), and anti-Kv4.3 antibodies ( C ). Blots were performed on starting material, P2 membrane fraction (P2), or on solubilized P2 proteins (sol) or after immunoprecipitation (IP1, IP2). Blots were probed with anti-SK3 (( A ), right panel), anti-Cav1.3 (( B ), right panel), anti-HCN4 (( A , B ) left panels; ( C ), right panel), anti-Kv4.3 (( C ), left panel), and anti-HCN2 (( C ), middle panel). P2: enriched P2 membrane fraction. IP1: immunoprecipitated proteins in CL47a buffer. IP2: immunoprecipitated proteins in CL48 buffer. sol: solubilisate. WB: Western blot (immunoblot).

    Techniques Used: Immunoprecipitation, Western Blot, Membrane

    Protein nano-environment of midbrain ion channels determined using affinity-purification mass spectrometry. ( A ) Left panel, Cav1.2 subunit ( Cacna1c gene) immunoprecipitations. Right panel, Cav1.3 subunit ( Cacna1d gene) immunoprecipitation. ( B ) SK3 subunit ( Kcnn3 gene) immunoprecipitation. ( C ) Left panel, HCN2 subunit immunoprecipitation. Right panel, HCN4 subunit immunoprecipitation. ( D ) Kv4.3 ( Kcnd3 gene) immunoprecipitation. High-confidence Class A (solid line) and low-confidence Class B (dashed lines) thresholds are displayed on the plots. Plain squares represent the bait and empty square interactants. The most relevant of the significant interactants are named.
    Figure Legend Snippet: Protein nano-environment of midbrain ion channels determined using affinity-purification mass spectrometry. ( A ) Left panel, Cav1.2 subunit ( Cacna1c gene) immunoprecipitations. Right panel, Cav1.3 subunit ( Cacna1d gene) immunoprecipitation. ( B ) SK3 subunit ( Kcnn3 gene) immunoprecipitation. ( C ) Left panel, HCN2 subunit immunoprecipitation. Right panel, HCN4 subunit immunoprecipitation. ( D ) Kv4.3 ( Kcnd3 gene) immunoprecipitation. High-confidence Class A (solid line) and low-confidence Class B (dashed lines) thresholds are displayed on the plots. Plain squares represent the bait and empty square interactants. The most relevant of the significant interactants are named.

    Techniques Used: Affinity Purification, Mass Spectrometry, Immunoprecipitation

    Visualization of the mass-spectrometry-based protein interaction network. The 6 ion channels of interest are depicted using different colors (blue, Kv4.3; pink and red, HCN2 and HCN4; yellow and orange, Cav1.2 and Cav1.3; green, SK3). Interactions are shown with lines of according colors. Thick lines represent Class A interactions, thinner lines Class B. Putative interactants were compared with the CRAPome 2.0 database (Contaminant Repository for Affinity Purification). Proteins identified in less than 1% of all CRAPome negative experiments were considered highly specific (black circles), between 1 and 2%, specific (grey circles), and between 2 and 10%, weakly specific (white circles) interactants.
    Figure Legend Snippet: Visualization of the mass-spectrometry-based protein interaction network. The 6 ion channels of interest are depicted using different colors (blue, Kv4.3; pink and red, HCN2 and HCN4; yellow and orange, Cav1.2 and Cav1.3; green, SK3). Interactions are shown with lines of according colors. Thick lines represent Class A interactions, thinner lines Class B. Putative interactants were compared with the CRAPome 2.0 database (Contaminant Repository for Affinity Purification). Proteins identified in less than 1% of all CRAPome negative experiments were considered highly specific (black circles), between 1 and 2%, specific (grey circles), and between 2 and 10%, weakly specific (white circles) interactants.

    Techniques Used: Mass Spectrometry, Affinity Purification

    HCN2, HCN4, and Sarm1 expression during midbrain postnatal development. Immunoblots of membrane-enriched fraction from adult (Ad) and 9 (P9) or 11 (P11) postnatal days Wt and Sarm1 −/− mice. Blots were probed with anti-HCN2 (left panel), anti-HCN4 (middle panel), or anti-Sarm1 (right panel) antibodies. WB: Western blot (immunoblot).
    Figure Legend Snippet: HCN2, HCN4, and Sarm1 expression during midbrain postnatal development. Immunoblots of membrane-enriched fraction from adult (Ad) and 9 (P9) or 11 (P11) postnatal days Wt and Sarm1 −/− mice. Blots were probed with anti-HCN2 (left panel), anti-HCN4 (middle panel), or anti-Sarm1 (right panel) antibodies. WB: Western blot (immunoblot).

    Techniques Used: Expressing, Western Blot, Membrane

    Cav1.3-HCN2-SK3 complex characterization in SNc DA neurons using proximity ligation assay. ( A ) Left, representative images of the PLA reaction (green) in wild-type (Wt) or Cav1.3 −/− SNc DA neurons using anti-Cav1.3 and anti-HCN2 antibodies. Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( B ) Representative images of the PLA reaction (green) in wild-type (Wt) SNc DA neurons using anti-SK3 and anti-HCN2 antibodies (top row) or anti-SK2 and anti-HCN2 antibodies (bottom row). Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( C ) Quantification of PLA fluorescent puncta on SNc DA neurons. Left, Cav1.3 and HCN2 PLA from 3 Wt (empty black circles; n = 21 images) or 3 Cav1.3 −/− (empty red circles; n = 18 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Middle, HCN2 and SK3 PLA (empty black circles; n = 11 images) or HCN2 and SK2 (empty orange circles; n = 7 images) from 2 independent experiments or anti-Kv4.3 and HCN2 (empty grey circle; n = 4 images) PLA from 1 experiment. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Right, Cav1.3 and SK3 PLA from 2 Wt (empty black circles; n = 10 images) or 2 Cav1.3 −/− (empty red circles; n = 8 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. ** p < 0.005; *** p < 0.001, Wilcoxon–Mann–Whitney test.
    Figure Legend Snippet: Cav1.3-HCN2-SK3 complex characterization in SNc DA neurons using proximity ligation assay. ( A ) Left, representative images of the PLA reaction (green) in wild-type (Wt) or Cav1.3 −/− SNc DA neurons using anti-Cav1.3 and anti-HCN2 antibodies. Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( B ) Representative images of the PLA reaction (green) in wild-type (Wt) SNc DA neurons using anti-SK3 and anti-HCN2 antibodies (top row) or anti-SK2 and anti-HCN2 antibodies (bottom row). Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( C ) Quantification of PLA fluorescent puncta on SNc DA neurons. Left, Cav1.3 and HCN2 PLA from 3 Wt (empty black circles; n = 21 images) or 3 Cav1.3 −/− (empty red circles; n = 18 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Middle, HCN2 and SK3 PLA (empty black circles; n = 11 images) or HCN2 and SK2 (empty orange circles; n = 7 images) from 2 independent experiments or anti-Kv4.3 and HCN2 (empty grey circle; n = 4 images) PLA from 1 experiment. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Right, Cav1.3 and SK3 PLA from 2 Wt (empty black circles; n = 10 images) or 2 Cav1.3 −/− (empty red circles; n = 8 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. ** p < 0.005; *** p < 0.001, Wilcoxon–Mann–Whitney test.

    Techniques Used: Proximity Ligation Assay, Immunolabeling, MANN-WHITNEY

    hcn2 antibody  (Alomone Labs)


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    Alomone Labs hcn2 antibody
    Shank3 deletion impairs the function and expression of <t>HCN2</t> channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).
    Hcn2 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hcn2 antibody/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hcn2 antibody - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice"

    Article Title: Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice

    Journal: Cell Reports Medicine

    doi: 10.1016/j.xcrm.2024.101534

    Shank3 deletion impairs the function and expression of HCN2 channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).
    Figure Legend Snippet: Shank3 deletion impairs the function and expression of HCN2 channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).

    Techniques Used: Expressing, Western Blot, Membrane, Two Tailed Test

    HCN2 overexpression in the VPM corrects electrophysiological alterations in Shank3 KO mice (A) Schematic of virus injection. (B) Representative traces of Ih currents. (C) Summary of Ih current density with different voltage steps, showing that the HCN2 overexpression (KO+HCN2) group displays increased Ih current density compared with the scramble (KO+Scr) group. (D) Quantification plot representing increased maximal Ih current density in the KO+HCN2 overexpression (OE) group. (C and D) n = 5 neurons from 3 mice for WT+Scr; n = 9 neurons from 3 mice for KO+HCN2; n = 11 from 3 mice for KO+Scr. (E) Representative images of immunoblotting, showing the HCN2 expression level. (F) Quantification of protein level, showing the effects of HCN2 OE in Shank3 KO mice. n = 3 mice for WT+Scr; n = 3 mice for KO+HCN2; n = 3 mice for KO+Scr. (G) Immunofluorescence images showing the HCN2 expression level in mCherry-positive (yellow arrows) and -negative cells (white arrows). Scale bar: 20 μm. (H) Quantification of fluorescence intensity in mCherry-positive and -negative cells. n = 127 mCherry-positive neurons and n = 136 mCherry-negative neurons from 3 mice. (I–M) Quantification of RMP (I), Rm (J), burst threshold (K), rebound burst threshold (L), and rebound burst latency (M), showing that HCN2 OE could rescue the intrinsic properties of VPM cells in Shank3 KO mice. n = 16 neurons from 3 mice for WT+Scr; n = 12 neurons from 3 mice for KO+HCN2; n = 19 from 3 mice for KO+Scr. (N) Schematic of virus injection and patch-clamp recordings. (O) Representative images showing EGFP and mCherry expression in the VPM. Scale bar: 10 μm. (P) Example traces of EPSPs with 40-Hz photoactivation. (Q) Quantification of the EPSP summation ratio, showing that VPM cells show reduced integration capability to cortical excitatory synaptic inputs with HCN2 OE in Shank3 KO mice. n = 7 neurons from 3 mice for WT+Scr; n = 6 neurons from 3 mice for KO+HCN2; n = 10 from 3 mice for KO+Scr. For statistical comparisons, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (C), one-way ANOVA (D, F, and I–M), two-tailed unpaired t test (H), and repeated-measurement ANOVA (Q). Detailed statistical information can be found in . Data are presented as mean ± SEM.
    Figure Legend Snippet: HCN2 overexpression in the VPM corrects electrophysiological alterations in Shank3 KO mice (A) Schematic of virus injection. (B) Representative traces of Ih currents. (C) Summary of Ih current density with different voltage steps, showing that the HCN2 overexpression (KO+HCN2) group displays increased Ih current density compared with the scramble (KO+Scr) group. (D) Quantification plot representing increased maximal Ih current density in the KO+HCN2 overexpression (OE) group. (C and D) n = 5 neurons from 3 mice for WT+Scr; n = 9 neurons from 3 mice for KO+HCN2; n = 11 from 3 mice for KO+Scr. (E) Representative images of immunoblotting, showing the HCN2 expression level. (F) Quantification of protein level, showing the effects of HCN2 OE in Shank3 KO mice. n = 3 mice for WT+Scr; n = 3 mice for KO+HCN2; n = 3 mice for KO+Scr. (G) Immunofluorescence images showing the HCN2 expression level in mCherry-positive (yellow arrows) and -negative cells (white arrows). Scale bar: 20 μm. (H) Quantification of fluorescence intensity in mCherry-positive and -negative cells. n = 127 mCherry-positive neurons and n = 136 mCherry-negative neurons from 3 mice. (I–M) Quantification of RMP (I), Rm (J), burst threshold (K), rebound burst threshold (L), and rebound burst latency (M), showing that HCN2 OE could rescue the intrinsic properties of VPM cells in Shank3 KO mice. n = 16 neurons from 3 mice for WT+Scr; n = 12 neurons from 3 mice for KO+HCN2; n = 19 from 3 mice for KO+Scr. (N) Schematic of virus injection and patch-clamp recordings. (O) Representative images showing EGFP and mCherry expression in the VPM. Scale bar: 10 μm. (P) Example traces of EPSPs with 40-Hz photoactivation. (Q) Quantification of the EPSP summation ratio, showing that VPM cells show reduced integration capability to cortical excitatory synaptic inputs with HCN2 OE in Shank3 KO mice. n = 7 neurons from 3 mice for WT+Scr; n = 6 neurons from 3 mice for KO+HCN2; n = 10 from 3 mice for KO+Scr. For statistical comparisons, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (C), one-way ANOVA (D, F, and I–M), two-tailed unpaired t test (H), and repeated-measurement ANOVA (Q). Detailed statistical information can be found in . Data are presented as mean ± SEM.

    Techniques Used: Over Expression, Virus, Injection, Western Blot, Expressing, Immunofluorescence, Fluorescence, Patch Clamp, Two Tailed Test


    Figure Legend Snippet:

    Techniques Used: Virus, Recombinant, Software

    hcn2  (Alomone Labs)


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    Alomone Labs hcn2
    Shank3 deletion impairs the function and expression of <t>HCN2</t> channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).
    Hcn2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hcn2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hcn2 - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice"

    Article Title: Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice

    Journal: Cell Reports Medicine

    doi: 10.1016/j.xcrm.2024.101534

    Shank3 deletion impairs the function and expression of HCN2 channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).
    Figure Legend Snippet: Shank3 deletion impairs the function and expression of HCN2 channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).

    Techniques Used: Expressing, Western Blot, Membrane, Two Tailed Test

    HCN2 overexpression in the VPM corrects electrophysiological alterations in Shank3 KO mice (A) Schematic of virus injection. (B) Representative traces of Ih currents. (C) Summary of Ih current density with different voltage steps, showing that the HCN2 overexpression (KO+HCN2) group displays increased Ih current density compared with the scramble (KO+Scr) group. (D) Quantification plot representing increased maximal Ih current density in the KO+HCN2 overexpression (OE) group. (C and D) n = 5 neurons from 3 mice for WT+Scr; n = 9 neurons from 3 mice for KO+HCN2; n = 11 from 3 mice for KO+Scr. (E) Representative images of immunoblotting, showing the HCN2 expression level. (F) Quantification of protein level, showing the effects of HCN2 OE in Shank3 KO mice. n = 3 mice for WT+Scr; n = 3 mice for KO+HCN2; n = 3 mice for KO+Scr. (G) Immunofluorescence images showing the HCN2 expression level in mCherry-positive (yellow arrows) and -negative cells (white arrows). Scale bar: 20 μm. (H) Quantification of fluorescence intensity in mCherry-positive and -negative cells. n = 127 mCherry-positive neurons and n = 136 mCherry-negative neurons from 3 mice. (I–M) Quantification of RMP (I), Rm (J), burst threshold (K), rebound burst threshold (L), and rebound burst latency (M), showing that HCN2 OE could rescue the intrinsic properties of VPM cells in Shank3 KO mice. n = 16 neurons from 3 mice for WT+Scr; n = 12 neurons from 3 mice for KO+HCN2; n = 19 from 3 mice for KO+Scr. (N) Schematic of virus injection and patch-clamp recordings. (O) Representative images showing EGFP and mCherry expression in the VPM. Scale bar: 10 μm. (P) Example traces of EPSPs with 40-Hz photoactivation. (Q) Quantification of the EPSP summation ratio, showing that VPM cells show reduced integration capability to cortical excitatory synaptic inputs with HCN2 OE in Shank3 KO mice. n = 7 neurons from 3 mice for WT+Scr; n = 6 neurons from 3 mice for KO+HCN2; n = 10 from 3 mice for KO+Scr. For statistical comparisons, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (C), one-way ANOVA (D, F, and I–M), two-tailed unpaired t test (H), and repeated-measurement ANOVA (Q). Detailed statistical information can be found in . Data are presented as mean ± SEM.
    Figure Legend Snippet: HCN2 overexpression in the VPM corrects electrophysiological alterations in Shank3 KO mice (A) Schematic of virus injection. (B) Representative traces of Ih currents. (C) Summary of Ih current density with different voltage steps, showing that the HCN2 overexpression (KO+HCN2) group displays increased Ih current density compared with the scramble (KO+Scr) group. (D) Quantification plot representing increased maximal Ih current density in the KO+HCN2 overexpression (OE) group. (C and D) n = 5 neurons from 3 mice for WT+Scr; n = 9 neurons from 3 mice for KO+HCN2; n = 11 from 3 mice for KO+Scr. (E) Representative images of immunoblotting, showing the HCN2 expression level. (F) Quantification of protein level, showing the effects of HCN2 OE in Shank3 KO mice. n = 3 mice for WT+Scr; n = 3 mice for KO+HCN2; n = 3 mice for KO+Scr. (G) Immunofluorescence images showing the HCN2 expression level in mCherry-positive (yellow arrows) and -negative cells (white arrows). Scale bar: 20 μm. (H) Quantification of fluorescence intensity in mCherry-positive and -negative cells. n = 127 mCherry-positive neurons and n = 136 mCherry-negative neurons from 3 mice. (I–M) Quantification of RMP (I), Rm (J), burst threshold (K), rebound burst threshold (L), and rebound burst latency (M), showing that HCN2 OE could rescue the intrinsic properties of VPM cells in Shank3 KO mice. n = 16 neurons from 3 mice for WT+Scr; n = 12 neurons from 3 mice for KO+HCN2; n = 19 from 3 mice for KO+Scr. (N) Schematic of virus injection and patch-clamp recordings. (O) Representative images showing EGFP and mCherry expression in the VPM. Scale bar: 10 μm. (P) Example traces of EPSPs with 40-Hz photoactivation. (Q) Quantification of the EPSP summation ratio, showing that VPM cells show reduced integration capability to cortical excitatory synaptic inputs with HCN2 OE in Shank3 KO mice. n = 7 neurons from 3 mice for WT+Scr; n = 6 neurons from 3 mice for KO+HCN2; n = 10 from 3 mice for KO+Scr. For statistical comparisons, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (C), one-way ANOVA (D, F, and I–M), two-tailed unpaired t test (H), and repeated-measurement ANOVA (Q). Detailed statistical information can be found in . Data are presented as mean ± SEM.

    Techniques Used: Over Expression, Virus, Injection, Western Blot, Expressing, Immunofluorescence, Fluorescence, Patch Clamp, Two Tailed Test


    Figure Legend Snippet:

    Techniques Used: Virus, Recombinant, Software

    rabbit anti hcn2  (Alomone Labs)


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    Alomone Labs rabbit anti hcn2
    Shank3 deletion impairs the function and expression of <t>HCN2</t> channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).
    Rabbit Anti Hcn2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    Images

    1) Product Images from "Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice"

    Article Title: Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice

    Journal: Cell Reports Medicine

    doi: 10.1016/j.xcrm.2024.101534

    Shank3 deletion impairs the function and expression of HCN2 channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).
    Figure Legend Snippet: Shank3 deletion impairs the function and expression of HCN2 channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).

    Techniques Used: Expressing, Western Blot, Membrane, Two Tailed Test

    HCN2 overexpression in the VPM corrects electrophysiological alterations in Shank3 KO mice (A) Schematic of virus injection. (B) Representative traces of Ih currents. (C) Summary of Ih current density with different voltage steps, showing that the HCN2 overexpression (KO+HCN2) group displays increased Ih current density compared with the scramble (KO+Scr) group. (D) Quantification plot representing increased maximal Ih current density in the KO+HCN2 overexpression (OE) group. (C and D) n = 5 neurons from 3 mice for WT+Scr; n = 9 neurons from 3 mice for KO+HCN2; n = 11 from 3 mice for KO+Scr. (E) Representative images of immunoblotting, showing the HCN2 expression level. (F) Quantification of protein level, showing the effects of HCN2 OE in Shank3 KO mice. n = 3 mice for WT+Scr; n = 3 mice for KO+HCN2; n = 3 mice for KO+Scr. (G) Immunofluorescence images showing the HCN2 expression level in mCherry-positive (yellow arrows) and -negative cells (white arrows). Scale bar: 20 μm. (H) Quantification of fluorescence intensity in mCherry-positive and -negative cells. n = 127 mCherry-positive neurons and n = 136 mCherry-negative neurons from 3 mice. (I–M) Quantification of RMP (I), Rm (J), burst threshold (K), rebound burst threshold (L), and rebound burst latency (M), showing that HCN2 OE could rescue the intrinsic properties of VPM cells in Shank3 KO mice. n = 16 neurons from 3 mice for WT+Scr; n = 12 neurons from 3 mice for KO+HCN2; n = 19 from 3 mice for KO+Scr. (N) Schematic of virus injection and patch-clamp recordings. (O) Representative images showing EGFP and mCherry expression in the VPM. Scale bar: 10 μm. (P) Example traces of EPSPs with 40-Hz photoactivation. (Q) Quantification of the EPSP summation ratio, showing that VPM cells show reduced integration capability to cortical excitatory synaptic inputs with HCN2 OE in Shank3 KO mice. n = 7 neurons from 3 mice for WT+Scr; n = 6 neurons from 3 mice for KO+HCN2; n = 10 from 3 mice for KO+Scr. For statistical comparisons, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (C), one-way ANOVA (D, F, and I–M), two-tailed unpaired t test (H), and repeated-measurement ANOVA (Q). Detailed statistical information can be found in . Data are presented as mean ± SEM.
    Figure Legend Snippet: HCN2 overexpression in the VPM corrects electrophysiological alterations in Shank3 KO mice (A) Schematic of virus injection. (B) Representative traces of Ih currents. (C) Summary of Ih current density with different voltage steps, showing that the HCN2 overexpression (KO+HCN2) group displays increased Ih current density compared with the scramble (KO+Scr) group. (D) Quantification plot representing increased maximal Ih current density in the KO+HCN2 overexpression (OE) group. (C and D) n = 5 neurons from 3 mice for WT+Scr; n = 9 neurons from 3 mice for KO+HCN2; n = 11 from 3 mice for KO+Scr. (E) Representative images of immunoblotting, showing the HCN2 expression level. (F) Quantification of protein level, showing the effects of HCN2 OE in Shank3 KO mice. n = 3 mice for WT+Scr; n = 3 mice for KO+HCN2; n = 3 mice for KO+Scr. (G) Immunofluorescence images showing the HCN2 expression level in mCherry-positive (yellow arrows) and -negative cells (white arrows). Scale bar: 20 μm. (H) Quantification of fluorescence intensity in mCherry-positive and -negative cells. n = 127 mCherry-positive neurons and n = 136 mCherry-negative neurons from 3 mice. (I–M) Quantification of RMP (I), Rm (J), burst threshold (K), rebound burst threshold (L), and rebound burst latency (M), showing that HCN2 OE could rescue the intrinsic properties of VPM cells in Shank3 KO mice. n = 16 neurons from 3 mice for WT+Scr; n = 12 neurons from 3 mice for KO+HCN2; n = 19 from 3 mice for KO+Scr. (N) Schematic of virus injection and patch-clamp recordings. (O) Representative images showing EGFP and mCherry expression in the VPM. Scale bar: 10 μm. (P) Example traces of EPSPs with 40-Hz photoactivation. (Q) Quantification of the EPSP summation ratio, showing that VPM cells show reduced integration capability to cortical excitatory synaptic inputs with HCN2 OE in Shank3 KO mice. n = 7 neurons from 3 mice for WT+Scr; n = 6 neurons from 3 mice for KO+HCN2; n = 10 from 3 mice for KO+Scr. For statistical comparisons, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (C), one-way ANOVA (D, F, and I–M), two-tailed unpaired t test (H), and repeated-measurement ANOVA (Q). Detailed statistical information can be found in . Data are presented as mean ± SEM.

    Techniques Used: Over Expression, Virus, Injection, Western Blot, Expressing, Immunofluorescence, Fluorescence, Patch Clamp, Two Tailed Test


    Figure Legend Snippet:

    Techniques Used: Virus, Recombinant, Software

    hcn2  (Alomone Labs)


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    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
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    Structured Review

    Alomone Labs hcn2
    Shank3 deletion impairs the function and expression of <t>HCN2</t> channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).
    Hcn2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hcn2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hcn2 - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice"

    Article Title: Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice

    Journal: Cell Reports Medicine

    doi: 10.1016/j.xcrm.2024.101534

    Shank3 deletion impairs the function and expression of HCN2 channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).
    Figure Legend Snippet: Shank3 deletion impairs the function and expression of HCN2 channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).

    Techniques Used: Expressing, Western Blot, Membrane, Two Tailed Test

    HCN2 overexpression in the VPM corrects electrophysiological alterations in Shank3 KO mice (A) Schematic of virus injection. (B) Representative traces of Ih currents. (C) Summary of Ih current density with different voltage steps, showing that the HCN2 overexpression (KO+HCN2) group displays increased Ih current density compared with the scramble (KO+Scr) group. (D) Quantification plot representing increased maximal Ih current density in the KO+HCN2 overexpression (OE) group. (C and D) n = 5 neurons from 3 mice for WT+Scr; n = 9 neurons from 3 mice for KO+HCN2; n = 11 from 3 mice for KO+Scr. (E) Representative images of immunoblotting, showing the HCN2 expression level. (F) Quantification of protein level, showing the effects of HCN2 OE in Shank3 KO mice. n = 3 mice for WT+Scr; n = 3 mice for KO+HCN2; n = 3 mice for KO+Scr. (G) Immunofluorescence images showing the HCN2 expression level in mCherry-positive (yellow arrows) and -negative cells (white arrows). Scale bar: 20 μm. (H) Quantification of fluorescence intensity in mCherry-positive and -negative cells. n = 127 mCherry-positive neurons and n = 136 mCherry-negative neurons from 3 mice. (I–M) Quantification of RMP (I), Rm (J), burst threshold (K), rebound burst threshold (L), and rebound burst latency (M), showing that HCN2 OE could rescue the intrinsic properties of VPM cells in Shank3 KO mice. n = 16 neurons from 3 mice for WT+Scr; n = 12 neurons from 3 mice for KO+HCN2; n = 19 from 3 mice for KO+Scr. (N) Schematic of virus injection and patch-clamp recordings. (O) Representative images showing EGFP and mCherry expression in the VPM. Scale bar: 10 μm. (P) Example traces of EPSPs with 40-Hz photoactivation. (Q) Quantification of the EPSP summation ratio, showing that VPM cells show reduced integration capability to cortical excitatory synaptic inputs with HCN2 OE in Shank3 KO mice. n = 7 neurons from 3 mice for WT+Scr; n = 6 neurons from 3 mice for KO+HCN2; n = 10 from 3 mice for KO+Scr. For statistical comparisons, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (C), one-way ANOVA (D, F, and I–M), two-tailed unpaired t test (H), and repeated-measurement ANOVA (Q). Detailed statistical information can be found in . Data are presented as mean ± SEM.
    Figure Legend Snippet: HCN2 overexpression in the VPM corrects electrophysiological alterations in Shank3 KO mice (A) Schematic of virus injection. (B) Representative traces of Ih currents. (C) Summary of Ih current density with different voltage steps, showing that the HCN2 overexpression (KO+HCN2) group displays increased Ih current density compared with the scramble (KO+Scr) group. (D) Quantification plot representing increased maximal Ih current density in the KO+HCN2 overexpression (OE) group. (C and D) n = 5 neurons from 3 mice for WT+Scr; n = 9 neurons from 3 mice for KO+HCN2; n = 11 from 3 mice for KO+Scr. (E) Representative images of immunoblotting, showing the HCN2 expression level. (F) Quantification of protein level, showing the effects of HCN2 OE in Shank3 KO mice. n = 3 mice for WT+Scr; n = 3 mice for KO+HCN2; n = 3 mice for KO+Scr. (G) Immunofluorescence images showing the HCN2 expression level in mCherry-positive (yellow arrows) and -negative cells (white arrows). Scale bar: 20 μm. (H) Quantification of fluorescence intensity in mCherry-positive and -negative cells. n = 127 mCherry-positive neurons and n = 136 mCherry-negative neurons from 3 mice. (I–M) Quantification of RMP (I), Rm (J), burst threshold (K), rebound burst threshold (L), and rebound burst latency (M), showing that HCN2 OE could rescue the intrinsic properties of VPM cells in Shank3 KO mice. n = 16 neurons from 3 mice for WT+Scr; n = 12 neurons from 3 mice for KO+HCN2; n = 19 from 3 mice for KO+Scr. (N) Schematic of virus injection and patch-clamp recordings. (O) Representative images showing EGFP and mCherry expression in the VPM. Scale bar: 10 μm. (P) Example traces of EPSPs with 40-Hz photoactivation. (Q) Quantification of the EPSP summation ratio, showing that VPM cells show reduced integration capability to cortical excitatory synaptic inputs with HCN2 OE in Shank3 KO mice. n = 7 neurons from 3 mice for WT+Scr; n = 6 neurons from 3 mice for KO+HCN2; n = 10 from 3 mice for KO+Scr. For statistical comparisons, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (C), one-way ANOVA (D, F, and I–M), two-tailed unpaired t test (H), and repeated-measurement ANOVA (Q). Detailed statistical information can be found in . Data are presented as mean ± SEM.

    Techniques Used: Over Expression, Virus, Injection, Western Blot, Expressing, Immunofluorescence, Fluorescence, Patch Clamp, Two Tailed Test


    Figure Legend Snippet:

    Techniques Used: Virus, Recombinant, Software

    hcn2  (Alomone Labs)


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    Alomone Labs hcn2
    Primary antibodies.
    Hcn2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs hcn2 antibody
    Shank3 deletion impairs the function and expression of <t>HCN2</t> channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).
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    Alomone Labs rabbit anti hcn2
    Shank3 deletion impairs the function and expression of <t>HCN2</t> channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).
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    Image Search Results


    Primary antibodies.

    Journal: Cells

    Article Title: High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons

    doi: 10.3390/cells13110944

    Figure Lengend Snippet: Primary antibodies.

    Article Snippet: IB , HCN2 , Rabbit , polyclonal , Alomone , APC-030 , AB_2313726 , 0.85.

    Techniques: Concentration Assay

    Characterization of the molecular interactions between Cav1.3, SK3, and HCN channels using co-immunoprecipitation and immunoblot. Proteins were membrane-extracted in CL47a solubilization buffer (IP1) or in CL48 buffer (IP2) and immunoprecipitated using anti-Cav1.3 ( A ), anti-SK3 ( B ), and anti-Kv4.3 antibodies ( C ). Blots were performed on starting material, P2 membrane fraction (P2), or on solubilized P2 proteins (sol) or after immunoprecipitation (IP1, IP2). Blots were probed with anti-SK3 (( A ), right panel), anti-Cav1.3 (( B ), right panel), anti-HCN4 (( A , B ) left panels; ( C ), right panel), anti-Kv4.3 (( C ), left panel), and anti-HCN2 (( C ), middle panel). P2: enriched P2 membrane fraction. IP1: immunoprecipitated proteins in CL47a buffer. IP2: immunoprecipitated proteins in CL48 buffer. sol: solubilisate. WB: Western blot (immunoblot).

    Journal: Cells

    Article Title: High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons

    doi: 10.3390/cells13110944

    Figure Lengend Snippet: Characterization of the molecular interactions between Cav1.3, SK3, and HCN channels using co-immunoprecipitation and immunoblot. Proteins were membrane-extracted in CL47a solubilization buffer (IP1) or in CL48 buffer (IP2) and immunoprecipitated using anti-Cav1.3 ( A ), anti-SK3 ( B ), and anti-Kv4.3 antibodies ( C ). Blots were performed on starting material, P2 membrane fraction (P2), or on solubilized P2 proteins (sol) or after immunoprecipitation (IP1, IP2). Blots were probed with anti-SK3 (( A ), right panel), anti-Cav1.3 (( B ), right panel), anti-HCN4 (( A , B ) left panels; ( C ), right panel), anti-Kv4.3 (( C ), left panel), and anti-HCN2 (( C ), middle panel). P2: enriched P2 membrane fraction. IP1: immunoprecipitated proteins in CL47a buffer. IP2: immunoprecipitated proteins in CL48 buffer. sol: solubilisate. WB: Western blot (immunoblot).

    Article Snippet: IB , HCN2 , Rabbit , polyclonal , Alomone , APC-030 , AB_2313726 , 0.85.

    Techniques: Immunoprecipitation, Western Blot, Membrane

    Protein nano-environment of midbrain ion channels determined using affinity-purification mass spectrometry. ( A ) Left panel, Cav1.2 subunit ( Cacna1c gene) immunoprecipitations. Right panel, Cav1.3 subunit ( Cacna1d gene) immunoprecipitation. ( B ) SK3 subunit ( Kcnn3 gene) immunoprecipitation. ( C ) Left panel, HCN2 subunit immunoprecipitation. Right panel, HCN4 subunit immunoprecipitation. ( D ) Kv4.3 ( Kcnd3 gene) immunoprecipitation. High-confidence Class A (solid line) and low-confidence Class B (dashed lines) thresholds are displayed on the plots. Plain squares represent the bait and empty square interactants. The most relevant of the significant interactants are named.

    Journal: Cells

    Article Title: High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons

    doi: 10.3390/cells13110944

    Figure Lengend Snippet: Protein nano-environment of midbrain ion channels determined using affinity-purification mass spectrometry. ( A ) Left panel, Cav1.2 subunit ( Cacna1c gene) immunoprecipitations. Right panel, Cav1.3 subunit ( Cacna1d gene) immunoprecipitation. ( B ) SK3 subunit ( Kcnn3 gene) immunoprecipitation. ( C ) Left panel, HCN2 subunit immunoprecipitation. Right panel, HCN4 subunit immunoprecipitation. ( D ) Kv4.3 ( Kcnd3 gene) immunoprecipitation. High-confidence Class A (solid line) and low-confidence Class B (dashed lines) thresholds are displayed on the plots. Plain squares represent the bait and empty square interactants. The most relevant of the significant interactants are named.

    Article Snippet: IB , HCN2 , Rabbit , polyclonal , Alomone , APC-030 , AB_2313726 , 0.85.

    Techniques: Affinity Purification, Mass Spectrometry, Immunoprecipitation

    Visualization of the mass-spectrometry-based protein interaction network. The 6 ion channels of interest are depicted using different colors (blue, Kv4.3; pink and red, HCN2 and HCN4; yellow and orange, Cav1.2 and Cav1.3; green, SK3). Interactions are shown with lines of according colors. Thick lines represent Class A interactions, thinner lines Class B. Putative interactants were compared with the CRAPome 2.0 database (Contaminant Repository for Affinity Purification). Proteins identified in less than 1% of all CRAPome negative experiments were considered highly specific (black circles), between 1 and 2%, specific (grey circles), and between 2 and 10%, weakly specific (white circles) interactants.

    Journal: Cells

    Article Title: High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons

    doi: 10.3390/cells13110944

    Figure Lengend Snippet: Visualization of the mass-spectrometry-based protein interaction network. The 6 ion channels of interest are depicted using different colors (blue, Kv4.3; pink and red, HCN2 and HCN4; yellow and orange, Cav1.2 and Cav1.3; green, SK3). Interactions are shown with lines of according colors. Thick lines represent Class A interactions, thinner lines Class B. Putative interactants were compared with the CRAPome 2.0 database (Contaminant Repository for Affinity Purification). Proteins identified in less than 1% of all CRAPome negative experiments were considered highly specific (black circles), between 1 and 2%, specific (grey circles), and between 2 and 10%, weakly specific (white circles) interactants.

    Article Snippet: IB , HCN2 , Rabbit , polyclonal , Alomone , APC-030 , AB_2313726 , 0.85.

    Techniques: Mass Spectrometry, Affinity Purification

    HCN2, HCN4, and Sarm1 expression during midbrain postnatal development. Immunoblots of membrane-enriched fraction from adult (Ad) and 9 (P9) or 11 (P11) postnatal days Wt and Sarm1 −/− mice. Blots were probed with anti-HCN2 (left panel), anti-HCN4 (middle panel), or anti-Sarm1 (right panel) antibodies. WB: Western blot (immunoblot).

    Journal: Cells

    Article Title: High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons

    doi: 10.3390/cells13110944

    Figure Lengend Snippet: HCN2, HCN4, and Sarm1 expression during midbrain postnatal development. Immunoblots of membrane-enriched fraction from adult (Ad) and 9 (P9) or 11 (P11) postnatal days Wt and Sarm1 −/− mice. Blots were probed with anti-HCN2 (left panel), anti-HCN4 (middle panel), or anti-Sarm1 (right panel) antibodies. WB: Western blot (immunoblot).

    Article Snippet: IB , HCN2 , Rabbit , polyclonal , Alomone , APC-030 , AB_2313726 , 0.85.

    Techniques: Expressing, Western Blot, Membrane

    Cav1.3-HCN2-SK3 complex characterization in SNc DA neurons using proximity ligation assay. ( A ) Left, representative images of the PLA reaction (green) in wild-type (Wt) or Cav1.3 −/− SNc DA neurons using anti-Cav1.3 and anti-HCN2 antibodies. Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( B ) Representative images of the PLA reaction (green) in wild-type (Wt) SNc DA neurons using anti-SK3 and anti-HCN2 antibodies (top row) or anti-SK2 and anti-HCN2 antibodies (bottom row). Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( C ) Quantification of PLA fluorescent puncta on SNc DA neurons. Left, Cav1.3 and HCN2 PLA from 3 Wt (empty black circles; n = 21 images) or 3 Cav1.3 −/− (empty red circles; n = 18 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Middle, HCN2 and SK3 PLA (empty black circles; n = 11 images) or HCN2 and SK2 (empty orange circles; n = 7 images) from 2 independent experiments or anti-Kv4.3 and HCN2 (empty grey circle; n = 4 images) PLA from 1 experiment. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Right, Cav1.3 and SK3 PLA from 2 Wt (empty black circles; n = 10 images) or 2 Cav1.3 −/− (empty red circles; n = 8 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. ** p < 0.005; *** p < 0.001, Wilcoxon–Mann–Whitney test.

    Journal: Cells

    Article Title: High-Resolution Proteomics Unravel a Native Functional Complex of Cav1.3, SK3, and Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels in Midbrain Dopaminergic Neurons

    doi: 10.3390/cells13110944

    Figure Lengend Snippet: Cav1.3-HCN2-SK3 complex characterization in SNc DA neurons using proximity ligation assay. ( A ) Left, representative images of the PLA reaction (green) in wild-type (Wt) or Cav1.3 −/− SNc DA neurons using anti-Cav1.3 and anti-HCN2 antibodies. Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( B ) Representative images of the PLA reaction (green) in wild-type (Wt) SNc DA neurons using anti-SK3 and anti-HCN2 antibodies (top row) or anti-SK2 and anti-HCN2 antibodies (bottom row). Middle, immunolabeling of SNc DA neurons with anti-TH antibodies (red). Right, merged images. Scale bars, 10 μm. ( C ) Quantification of PLA fluorescent puncta on SNc DA neurons. Left, Cav1.3 and HCN2 PLA from 3 Wt (empty black circles; n = 21 images) or 3 Cav1.3 −/− (empty red circles; n = 18 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Middle, HCN2 and SK3 PLA (empty black circles; n = 11 images) or HCN2 and SK2 (empty orange circles; n = 7 images) from 2 independent experiments or anti-Kv4.3 and HCN2 (empty grey circle; n = 4 images) PLA from 1 experiment. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. Right, Cav1.3 and SK3 PLA from 2 Wt (empty black circles; n = 10 images) or 2 Cav1.3 −/− (empty red circles; n = 8 images) mice. Superimposed box and whiskers plots allow us to visualize the median and interquartile range. ** p < 0.005; *** p < 0.001, Wilcoxon–Mann–Whitney test.

    Article Snippet: IB , HCN2 , Rabbit , polyclonal , Alomone , APC-030 , AB_2313726 , 0.85.

    Techniques: Proximity Ligation Assay, Immunolabeling, MANN-WHITNEY

    Shank3 deletion impairs the function and expression of HCN2 channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).

    Journal: Cell Reports Medicine

    Article Title: Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice

    doi: 10.1016/j.xcrm.2024.101534

    Figure Lengend Snippet: Shank3 deletion impairs the function and expression of HCN2 channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).

    Article Snippet: The sections were incubated overnight at room temperature with Rabbit anti HCN2 antibody (1:200; Alomone labs, APC-030).

    Techniques: Expressing, Western Blot, Membrane, Two Tailed Test

    HCN2 overexpression in the VPM corrects electrophysiological alterations in Shank3 KO mice (A) Schematic of virus injection. (B) Representative traces of Ih currents. (C) Summary of Ih current density with different voltage steps, showing that the HCN2 overexpression (KO+HCN2) group displays increased Ih current density compared with the scramble (KO+Scr) group. (D) Quantification plot representing increased maximal Ih current density in the KO+HCN2 overexpression (OE) group. (C and D) n = 5 neurons from 3 mice for WT+Scr; n = 9 neurons from 3 mice for KO+HCN2; n = 11 from 3 mice for KO+Scr. (E) Representative images of immunoblotting, showing the HCN2 expression level. (F) Quantification of protein level, showing the effects of HCN2 OE in Shank3 KO mice. n = 3 mice for WT+Scr; n = 3 mice for KO+HCN2; n = 3 mice for KO+Scr. (G) Immunofluorescence images showing the HCN2 expression level in mCherry-positive (yellow arrows) and -negative cells (white arrows). Scale bar: 20 μm. (H) Quantification of fluorescence intensity in mCherry-positive and -negative cells. n = 127 mCherry-positive neurons and n = 136 mCherry-negative neurons from 3 mice. (I–M) Quantification of RMP (I), Rm (J), burst threshold (K), rebound burst threshold (L), and rebound burst latency (M), showing that HCN2 OE could rescue the intrinsic properties of VPM cells in Shank3 KO mice. n = 16 neurons from 3 mice for WT+Scr; n = 12 neurons from 3 mice for KO+HCN2; n = 19 from 3 mice for KO+Scr. (N) Schematic of virus injection and patch-clamp recordings. (O) Representative images showing EGFP and mCherry expression in the VPM. Scale bar: 10 μm. (P) Example traces of EPSPs with 40-Hz photoactivation. (Q) Quantification of the EPSP summation ratio, showing that VPM cells show reduced integration capability to cortical excitatory synaptic inputs with HCN2 OE in Shank3 KO mice. n = 7 neurons from 3 mice for WT+Scr; n = 6 neurons from 3 mice for KO+HCN2; n = 10 from 3 mice for KO+Scr. For statistical comparisons, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (C), one-way ANOVA (D, F, and I–M), two-tailed unpaired t test (H), and repeated-measurement ANOVA (Q). Detailed statistical information can be found in . Data are presented as mean ± SEM.

    Journal: Cell Reports Medicine

    Article Title: Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice

    doi: 10.1016/j.xcrm.2024.101534

    Figure Lengend Snippet: HCN2 overexpression in the VPM corrects electrophysiological alterations in Shank3 KO mice (A) Schematic of virus injection. (B) Representative traces of Ih currents. (C) Summary of Ih current density with different voltage steps, showing that the HCN2 overexpression (KO+HCN2) group displays increased Ih current density compared with the scramble (KO+Scr) group. (D) Quantification plot representing increased maximal Ih current density in the KO+HCN2 overexpression (OE) group. (C and D) n = 5 neurons from 3 mice for WT+Scr; n = 9 neurons from 3 mice for KO+HCN2; n = 11 from 3 mice for KO+Scr. (E) Representative images of immunoblotting, showing the HCN2 expression level. (F) Quantification of protein level, showing the effects of HCN2 OE in Shank3 KO mice. n = 3 mice for WT+Scr; n = 3 mice for KO+HCN2; n = 3 mice for KO+Scr. (G) Immunofluorescence images showing the HCN2 expression level in mCherry-positive (yellow arrows) and -negative cells (white arrows). Scale bar: 20 μm. (H) Quantification of fluorescence intensity in mCherry-positive and -negative cells. n = 127 mCherry-positive neurons and n = 136 mCherry-negative neurons from 3 mice. (I–M) Quantification of RMP (I), Rm (J), burst threshold (K), rebound burst threshold (L), and rebound burst latency (M), showing that HCN2 OE could rescue the intrinsic properties of VPM cells in Shank3 KO mice. n = 16 neurons from 3 mice for WT+Scr; n = 12 neurons from 3 mice for KO+HCN2; n = 19 from 3 mice for KO+Scr. (N) Schematic of virus injection and patch-clamp recordings. (O) Representative images showing EGFP and mCherry expression in the VPM. Scale bar: 10 μm. (P) Example traces of EPSPs with 40-Hz photoactivation. (Q) Quantification of the EPSP summation ratio, showing that VPM cells show reduced integration capability to cortical excitatory synaptic inputs with HCN2 OE in Shank3 KO mice. n = 7 neurons from 3 mice for WT+Scr; n = 6 neurons from 3 mice for KO+HCN2; n = 10 from 3 mice for KO+Scr. For statistical comparisons, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (C), one-way ANOVA (D, F, and I–M), two-tailed unpaired t test (H), and repeated-measurement ANOVA (Q). Detailed statistical information can be found in . Data are presented as mean ± SEM.

    Article Snippet: The sections were incubated overnight at room temperature with Rabbit anti HCN2 antibody (1:200; Alomone labs, APC-030).

    Techniques: Over Expression, Virus, Injection, Western Blot, Expressing, Immunofluorescence, Fluorescence, Patch Clamp, Two Tailed Test

    Journal: Cell Reports Medicine

    Article Title: Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice

    doi: 10.1016/j.xcrm.2024.101534

    Figure Lengend Snippet:

    Article Snippet: The sections were incubated overnight at room temperature with Rabbit anti HCN2 antibody (1:200; Alomone labs, APC-030).

    Techniques: Virus, Recombinant, Software

    Shank3 deletion impairs the function and expression of HCN2 channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).

    Journal: Cell Reports Medicine

    Article Title: Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice

    doi: 10.1016/j.xcrm.2024.101534

    Figure Lengend Snippet: Shank3 deletion impairs the function and expression of HCN2 channels in the VPM (A) Representative traces of Ih currents in Shank3 WT (left) and KO (right) cells. (B) Summary of Ih current density with different voltage steps, showing decreased HCN mediated current density in Shank3 KO cells. (C) Quantification plot representing reduced maximal Ih current density in Shank3 KO cells. (B and C) n = 9 neurons from 3 mice for WT; n = 9 neurons from 3 mice for KO. (D) Representative images of immunoblotting for HCN1, HCN2, and HCN4 of the VPM total protein in Shank3 WT and KO. (E) Quantification plot showing that the expression level of HCN2 is decreased in Shank3 KO mice. (F) Representative images of immunoblotting for HCN2 of the VPM synaptoneurosomal protein in Shank3 WT and KO. (G) Quantification plot showing that the expression level of HCN2 is reduced in Shank3 KO mice. (E and G) n = 3 mice for WT; n = 3 mice for KO. (H) Representative immunogold micrographs showing the subcellular location of HCN2 (red arrows point to HCN2 in the membrane) in somatic (left) and dendritic regions (right) in Shank3 WT (top) and KO (bottom). Scale bar: 0.5 μm. (I) Density of HCN2-positive particles in somata (left) and dendrites (right). (J) Linear density of HCN2-positive particles in the neuronal membranes in somata (left) and dendrites (right). (I and J) n = 20 somata and 99 dendrites from 3 mice for WT; n = 16 somata and 95 dendrites from 3 mice for KO. For statistical comparisons, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (B) and two-tailed unpaired t test (C, E, G, I, and J). Detailed statistical information can be found in . Data are presented as mean ± SEM (B, C, E, and G) and median ± 95% CI (I and J).

    Article Snippet: Rabbit anti HCN2 , Alomone labs , Cat. # APC-030; RRID: AB_2313726.

    Techniques: Expressing, Western Blot, Membrane, Two Tailed Test

    HCN2 overexpression in the VPM corrects electrophysiological alterations in Shank3 KO mice (A) Schematic of virus injection. (B) Representative traces of Ih currents. (C) Summary of Ih current density with different voltage steps, showing that the HCN2 overexpression (KO+HCN2) group displays increased Ih current density compared with the scramble (KO+Scr) group. (D) Quantification plot representing increased maximal Ih current density in the KO+HCN2 overexpression (OE) group. (C and D) n = 5 neurons from 3 mice for WT+Scr; n = 9 neurons from 3 mice for KO+HCN2; n = 11 from 3 mice for KO+Scr. (E) Representative images of immunoblotting, showing the HCN2 expression level. (F) Quantification of protein level, showing the effects of HCN2 OE in Shank3 KO mice. n = 3 mice for WT+Scr; n = 3 mice for KO+HCN2; n = 3 mice for KO+Scr. (G) Immunofluorescence images showing the HCN2 expression level in mCherry-positive (yellow arrows) and -negative cells (white arrows). Scale bar: 20 μm. (H) Quantification of fluorescence intensity in mCherry-positive and -negative cells. n = 127 mCherry-positive neurons and n = 136 mCherry-negative neurons from 3 mice. (I–M) Quantification of RMP (I), Rm (J), burst threshold (K), rebound burst threshold (L), and rebound burst latency (M), showing that HCN2 OE could rescue the intrinsic properties of VPM cells in Shank3 KO mice. n = 16 neurons from 3 mice for WT+Scr; n = 12 neurons from 3 mice for KO+HCN2; n = 19 from 3 mice for KO+Scr. (N) Schematic of virus injection and patch-clamp recordings. (O) Representative images showing EGFP and mCherry expression in the VPM. Scale bar: 10 μm. (P) Example traces of EPSPs with 40-Hz photoactivation. (Q) Quantification of the EPSP summation ratio, showing that VPM cells show reduced integration capability to cortical excitatory synaptic inputs with HCN2 OE in Shank3 KO mice. n = 7 neurons from 3 mice for WT+Scr; n = 6 neurons from 3 mice for KO+HCN2; n = 10 from 3 mice for KO+Scr. For statistical comparisons, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (C), one-way ANOVA (D, F, and I–M), two-tailed unpaired t test (H), and repeated-measurement ANOVA (Q). Detailed statistical information can be found in . Data are presented as mean ± SEM.

    Journal: Cell Reports Medicine

    Article Title: Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice

    doi: 10.1016/j.xcrm.2024.101534

    Figure Lengend Snippet: HCN2 overexpression in the VPM corrects electrophysiological alterations in Shank3 KO mice (A) Schematic of virus injection. (B) Representative traces of Ih currents. (C) Summary of Ih current density with different voltage steps, showing that the HCN2 overexpression (KO+HCN2) group displays increased Ih current density compared with the scramble (KO+Scr) group. (D) Quantification plot representing increased maximal Ih current density in the KO+HCN2 overexpression (OE) group. (C and D) n = 5 neurons from 3 mice for WT+Scr; n = 9 neurons from 3 mice for KO+HCN2; n = 11 from 3 mice for KO+Scr. (E) Representative images of immunoblotting, showing the HCN2 expression level. (F) Quantification of protein level, showing the effects of HCN2 OE in Shank3 KO mice. n = 3 mice for WT+Scr; n = 3 mice for KO+HCN2; n = 3 mice for KO+Scr. (G) Immunofluorescence images showing the HCN2 expression level in mCherry-positive (yellow arrows) and -negative cells (white arrows). Scale bar: 20 μm. (H) Quantification of fluorescence intensity in mCherry-positive and -negative cells. n = 127 mCherry-positive neurons and n = 136 mCherry-negative neurons from 3 mice. (I–M) Quantification of RMP (I), Rm (J), burst threshold (K), rebound burst threshold (L), and rebound burst latency (M), showing that HCN2 OE could rescue the intrinsic properties of VPM cells in Shank3 KO mice. n = 16 neurons from 3 mice for WT+Scr; n = 12 neurons from 3 mice for KO+HCN2; n = 19 from 3 mice for KO+Scr. (N) Schematic of virus injection and patch-clamp recordings. (O) Representative images showing EGFP and mCherry expression in the VPM. Scale bar: 10 μm. (P) Example traces of EPSPs with 40-Hz photoactivation. (Q) Quantification of the EPSP summation ratio, showing that VPM cells show reduced integration capability to cortical excitatory synaptic inputs with HCN2 OE in Shank3 KO mice. n = 7 neurons from 3 mice for WT+Scr; n = 6 neurons from 3 mice for KO+HCN2; n = 10 from 3 mice for KO+Scr. For statistical comparisons, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Friedman’s M test (C), one-way ANOVA (D, F, and I–M), two-tailed unpaired t test (H), and repeated-measurement ANOVA (Q). Detailed statistical information can be found in . Data are presented as mean ± SEM.

    Article Snippet: Rabbit anti HCN2 , Alomone labs , Cat. # APC-030; RRID: AB_2313726.

    Techniques: Over Expression, Virus, Injection, Western Blot, Expressing, Immunofluorescence, Fluorescence, Patch Clamp, Two Tailed Test

    Journal: Cell Reports Medicine

    Article Title: Restoring thalamocortical circuit dysfunction by correcting HCN channelopathy in Shank3 mutant mice

    doi: 10.1016/j.xcrm.2024.101534

    Figure Lengend Snippet:

    Article Snippet: Rabbit anti HCN2 , Alomone labs , Cat. # APC-030; RRID: AB_2313726.

    Techniques: Virus, Recombinant, Software