ca v 1 2  (Alomone Labs)


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    Alomone Labs ca v 1 2
    Primary antibodies.
    Ca V 1 2, 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/ca v 1 2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ca v 1 2 - 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


    Figure Legend Snippet: Primary antibodies.

    Techniques Used: Concentration Assay

    rabbit polyclonal igg anti ca v 1 2  (Alomone Labs)


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    Alomone Labs rabbit polyclonal igg anti ca v 1 2
    a Single-molecule localization microscopy (SMLM) map showing Ca V 1.2 channel localization and distribution in the t-tubules of young and old ventricular myocytes with or without ISO stimulation. Yellow boxes indicate the location of the regions of interest magnified in the top right of each image. b Dot-plots showing mean Ca V 1.2 channel cluster areas in young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 4, n = 11; ISO: N = 4, n = 10) myocytes. c , d show the same for RyR2 in young (control: N = 3, n = 15; ISO: N = 3, n = 15) and old (control: N = 3, n = 12; ISO: N = 3, n = 11) myocytes. Statistical analyses on data summarized in ( b , d ) were performed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in b and d are from NIA young. Data are presented as mean ± SEM. Source data are provided in the Source Data file.
    Rabbit Polyclonal Igg Anti Ca V 1 2, 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/rabbit polyclonal igg anti ca v 1 2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal igg anti ca v 1 2 - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "BIN1 knockdown rescues systolic dysfunction in aging male mouse hearts"

    Article Title: BIN1 knockdown rescues systolic dysfunction in aging male mouse hearts

    Journal: Nature Communications

    doi: 10.1038/s41467-024-47847-8

    a Single-molecule localization microscopy (SMLM) map showing Ca V 1.2 channel localization and distribution in the t-tubules of young and old ventricular myocytes with or without ISO stimulation. Yellow boxes indicate the location of the regions of interest magnified in the top right of each image. b Dot-plots showing mean Ca V 1.2 channel cluster areas in young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 4, n = 11; ISO: N = 4, n = 10) myocytes. c , d show the same for RyR2 in young (control: N = 3, n = 15; ISO: N = 3, n = 15) and old (control: N = 3, n = 12; ISO: N = 3, n = 11) myocytes. Statistical analyses on data summarized in ( b , d ) were performed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in b and d are from NIA young. Data are presented as mean ± SEM. Source data are provided in the Source Data file.
    Figure Legend Snippet: a Single-molecule localization microscopy (SMLM) map showing Ca V 1.2 channel localization and distribution in the t-tubules of young and old ventricular myocytes with or without ISO stimulation. Yellow boxes indicate the location of the regions of interest magnified in the top right of each image. b Dot-plots showing mean Ca V 1.2 channel cluster areas in young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 4, n = 11; ISO: N = 4, n = 10) myocytes. c , d show the same for RyR2 in young (control: N = 3, n = 15; ISO: N = 3, n = 15) and old (control: N = 3, n = 12; ISO: N = 3, n = 11) myocytes. Statistical analyses on data summarized in ( b , d ) were performed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in b and d are from NIA young. Data are presented as mean ± SEM. Source data are provided in the Source Data file.

    Techniques Used: Microscopy, Comparison

    a Airyscan super-resolution images of Ca V 1.2 (green) and EEA1 (magenta) immunostained myocytes with and without ISO. Bottom: Binary colocalization maps show pixels in which Ca V 1.2 and EEA1 completely overlapped. b dot-plots summarizing % colocalization between EEA1 and Ca V 1.2 young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 3, n = 14; ISO: N = 3, n = 14) myocytes, and c EEA1-positive endosome areas in young (control: N = 3, n = 15; ISO: N = 3, n = 16) and old (control: N = 3, n = 14; ISO: N = 3, n = 13) myocytes. Data were analyzed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in ( b , c ) are from JAX young. Note no significant differences in EEA1/Ca V 1.2 colocalization, responsivity to ISO, or endosome size was detected when young JAX and young NIA myocytes were compared (Supplementary Fig.  ). Data are presented as mean ± SEM. Source data are provided in the Source Data file.
    Figure Legend Snippet: a Airyscan super-resolution images of Ca V 1.2 (green) and EEA1 (magenta) immunostained myocytes with and without ISO. Bottom: Binary colocalization maps show pixels in which Ca V 1.2 and EEA1 completely overlapped. b dot-plots summarizing % colocalization between EEA1 and Ca V 1.2 young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 3, n = 14; ISO: N = 3, n = 14) myocytes, and c EEA1-positive endosome areas in young (control: N = 3, n = 15; ISO: N = 3, n = 16) and old (control: N = 3, n = 14; ISO: N = 3, n = 13) myocytes. Data were analyzed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in ( b , c ) are from JAX young. Note no significant differences in EEA1/Ca V 1.2 colocalization, responsivity to ISO, or endosome size was detected when young JAX and young NIA myocytes were compared (Supplementary Fig. ). Data are presented as mean ± SEM. Source data are provided in the Source Data file.

    Techniques Used: Comparison

    a Representative whole-cell currents from shRNA-scrmb and shRNA-mBIN1 myocytes before and during ISO application. b fold change in peak I Ca with ISO in young, old, shRNA-scrmb ( N = 3, n = 7), and shRNA-mBIN1 ( N = 3, n = 9) myocytes. c Representative Ca 2+ transients recorded from old shRNA-scrmb and shRNA-mBIN1 myocytes before and after ISO.  Fold increase after ISO from young, old, shRNA-scrmb ( N = 3, n = 14), and shRNA-mBIN1 ( N = 5, n = 18) myocytes. e SMLM localization maps showing Ca V 1.2 channel localization on t-tubules of myocytes from old shRNA-scrmb and shRNA-mBIN1, with or without ISO stimulation. Regions of interest are highlighted by yellow boxes. f Fold change in mean Ca V 1.2 channel cluster area with ISO in the young, old, old shRNA-scrmb (control: N = 3, n = 14; ISO: N = 3, n = 13) and shRNA-mBIN1 myocytes (control: N = 3, n = 12; ISO: N = 3, n = 11). g , h show the same layout for RyR2 immunostained old shRNA-scrmb (control: N = 3, n = 12; ISO: N = 3, n = 13) and shRNA-mBIN1 myocytes (control: N = 3, n = 9; ISO: N = 3, n = 8). Old and young data points in ( b ,  , f , h ) are reproduced from data in Figs.  b, h, and  b,  respectively. Statistical analysis was performed on data in ( b ,  , f , h ) using one-way ANOVAs with multiple comparison post-hoc tests. Young data in ( b ) are pooled from NIA young and JAX young myocytes, data in (  , f , h ) are from NIA young myocytes. Note there was no significant difference in I Ca , Ca V 1.2, and RyR2 cluster areas when JAX and NIA young mice were compared (see Supplementary Fig.  ). Data are presented as mean ± SEM. Source data are provided in the Source Data file.
    Figure Legend Snippet: a Representative whole-cell currents from shRNA-scrmb and shRNA-mBIN1 myocytes before and during ISO application. b fold change in peak I Ca with ISO in young, old, shRNA-scrmb ( N = 3, n = 7), and shRNA-mBIN1 ( N = 3, n = 9) myocytes. c Representative Ca 2+ transients recorded from old shRNA-scrmb and shRNA-mBIN1 myocytes before and after ISO. Fold increase after ISO from young, old, shRNA-scrmb ( N = 3, n = 14), and shRNA-mBIN1 ( N = 5, n = 18) myocytes. e SMLM localization maps showing Ca V 1.2 channel localization on t-tubules of myocytes from old shRNA-scrmb and shRNA-mBIN1, with or without ISO stimulation. Regions of interest are highlighted by yellow boxes. f Fold change in mean Ca V 1.2 channel cluster area with ISO in the young, old, old shRNA-scrmb (control: N = 3, n = 14; ISO: N = 3, n = 13) and shRNA-mBIN1 myocytes (control: N = 3, n = 12; ISO: N = 3, n = 11). g , h show the same layout for RyR2 immunostained old shRNA-scrmb (control: N = 3, n = 12; ISO: N = 3, n = 13) and shRNA-mBIN1 myocytes (control: N = 3, n = 9; ISO: N = 3, n = 8). Old and young data points in ( b , , f , h ) are reproduced from data in Figs. b, h, and b, respectively. Statistical analysis was performed on data in ( b , , f , h ) using one-way ANOVAs with multiple comparison post-hoc tests. Young data in ( b ) are pooled from NIA young and JAX young myocytes, data in ( , f , h ) are from NIA young myocytes. Note there was no significant difference in I Ca , Ca V 1.2, and RyR2 cluster areas when JAX and NIA young mice were compared (see Supplementary Fig. ). Data are presented as mean ± SEM. Source data are provided in the Source Data file.

    Techniques Used: shRNA, Comparison

    The main findings of our study graphically illustrated and summarized. Top : In healthy young cells, Ca V 1.2 channels undergo endosomal recycling, where channels on endosomes are either marked for degradation through the late endosome pathway or are recycled to the sarcolemma through the fast and slow recycling pathways. Following β -adrenergic receptor ( β -AR) stimulation, a pool of channels localized to endosomes are mobilized to the membrane, resulting in larger Ca V 1.2 clusters along t-tubules. Across the dyad, RyR2 clusters on the sarcoplasmic reticulum also increase following βAR stimulation ensuring efficient Ca 2+ -induced Ca 2+ -release. This increase in cytosolic Ca 2+ , along with increased phosphorylation of cardiac Troponin I (cTnI) and cardiac myosin binding protein-C (cMyBP-C) within the sarcomere, allows for enhanced contractility under acute stress to cope with elevated hemodynamic and metabolic demands. Bottom : In aging, Bridging Integrator 1 (BIN1) protein levels are increased, accompanied by a swelling of endosomes and subsequent dysregulation of endosomal trafficking of Ca V 1.2. Ca V 1.2 and RyR2 channels are basally super-clustered at the dyads and lose β -AR responsivity. Reduced phosphorylation of cTnI and cMyBP-C result in systolic and diastolic dysfunction. BIN1 knockdown in aging recovers RyR2 clustering plasticity and Ca 2+ transient responsivity to β- AR stimulation. Phosphorylation of cMyBP-C is basally restored, and contractility is recovered to youthful levels. Thus, BIN1 knockdown rejuvenates the aging heart. Created with BioRender.com.
    Figure Legend Snippet: The main findings of our study graphically illustrated and summarized. Top : In healthy young cells, Ca V 1.2 channels undergo endosomal recycling, where channels on endosomes are either marked for degradation through the late endosome pathway or are recycled to the sarcolemma through the fast and slow recycling pathways. Following β -adrenergic receptor ( β -AR) stimulation, a pool of channels localized to endosomes are mobilized to the membrane, resulting in larger Ca V 1.2 clusters along t-tubules. Across the dyad, RyR2 clusters on the sarcoplasmic reticulum also increase following βAR stimulation ensuring efficient Ca 2+ -induced Ca 2+ -release. This increase in cytosolic Ca 2+ , along with increased phosphorylation of cardiac Troponin I (cTnI) and cardiac myosin binding protein-C (cMyBP-C) within the sarcomere, allows for enhanced contractility under acute stress to cope with elevated hemodynamic and metabolic demands. Bottom : In aging, Bridging Integrator 1 (BIN1) protein levels are increased, accompanied by a swelling of endosomes and subsequent dysregulation of endosomal trafficking of Ca V 1.2. Ca V 1.2 and RyR2 channels are basally super-clustered at the dyads and lose β -AR responsivity. Reduced phosphorylation of cTnI and cMyBP-C result in systolic and diastolic dysfunction. BIN1 knockdown in aging recovers RyR2 clustering plasticity and Ca 2+ transient responsivity to β- AR stimulation. Phosphorylation of cMyBP-C is basally restored, and contractility is recovered to youthful levels. Thus, BIN1 knockdown rejuvenates the aging heart. Created with BioRender.com.

    Techniques Used: Membrane, Binding Assay

    rabbit polyclonal igg anti ca v 1 2  (Alomone Labs)


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    Alomone Labs rabbit polyclonal igg anti ca v 1 2
    a Single-molecule localization microscopy (SMLM) map showing Ca V 1.2 channel localization and distribution in the t-tubules of young and old ventricular myocytes with or without ISO stimulation. Yellow boxes indicate the location of the regions of interest magnified in the top right of each image. b Dot-plots showing mean Ca V 1.2 channel cluster areas in young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 4, n = 11; ISO: N = 4, n = 10) myocytes. c , d show the same for RyR2 in young (control: N = 3, n = 15; ISO: N = 3, n = 15) and old (control: N = 3, n = 12; ISO: N = 3, n = 11) myocytes. Statistical analyses on data summarized in ( b , d ) were performed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in b and d are from NIA young. Data are presented as mean ± SEM. Source data are provided in the Source Data file.
    Rabbit Polyclonal Igg Anti Ca V 1 2, 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/rabbit polyclonal igg anti ca v 1 2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal igg anti ca v 1 2 - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "BIN1 knockdown rescues systolic dysfunction in aging male mouse hearts"

    Article Title: BIN1 knockdown rescues systolic dysfunction in aging male mouse hearts

    Journal: Nature Communications

    doi: 10.1038/s41467-024-47847-8

    a Single-molecule localization microscopy (SMLM) map showing Ca V 1.2 channel localization and distribution in the t-tubules of young and old ventricular myocytes with or without ISO stimulation. Yellow boxes indicate the location of the regions of interest magnified in the top right of each image. b Dot-plots showing mean Ca V 1.2 channel cluster areas in young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 4, n = 11; ISO: N = 4, n = 10) myocytes. c , d show the same for RyR2 in young (control: N = 3, n = 15; ISO: N = 3, n = 15) and old (control: N = 3, n = 12; ISO: N = 3, n = 11) myocytes. Statistical analyses on data summarized in ( b , d ) were performed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in b and d are from NIA young. Data are presented as mean ± SEM. Source data are provided in the Source Data file.
    Figure Legend Snippet: a Single-molecule localization microscopy (SMLM) map showing Ca V 1.2 channel localization and distribution in the t-tubules of young and old ventricular myocytes with or without ISO stimulation. Yellow boxes indicate the location of the regions of interest magnified in the top right of each image. b Dot-plots showing mean Ca V 1.2 channel cluster areas in young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 4, n = 11; ISO: N = 4, n = 10) myocytes. c , d show the same for RyR2 in young (control: N = 3, n = 15; ISO: N = 3, n = 15) and old (control: N = 3, n = 12; ISO: N = 3, n = 11) myocytes. Statistical analyses on data summarized in ( b , d ) were performed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in b and d are from NIA young. Data are presented as mean ± SEM. Source data are provided in the Source Data file.

    Techniques Used: Microscopy, Comparison

    a Airyscan super-resolution images of Ca V 1.2 (green) and EEA1 (magenta) immunostained myocytes with and without ISO. Bottom: Binary colocalization maps show pixels in which Ca V 1.2 and EEA1 completely overlapped. b dot-plots summarizing % colocalization between EEA1 and Ca V 1.2 young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 3, n = 14; ISO: N = 3, n = 14) myocytes, and c EEA1-positive endosome areas in young (control: N = 3, n = 15; ISO: N = 3, n = 16) and old (control: N = 3, n = 14; ISO: N = 3, n = 13) myocytes. Data were analyzed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in ( b , c ) are from JAX young. Note no significant differences in EEA1/Ca V 1.2 colocalization, responsivity to ISO, or endosome size was detected when young JAX and young NIA myocytes were compared (Supplementary Fig.  ). Data are presented as mean ± SEM. Source data are provided in the Source Data file.
    Figure Legend Snippet: a Airyscan super-resolution images of Ca V 1.2 (green) and EEA1 (magenta) immunostained myocytes with and without ISO. Bottom: Binary colocalization maps show pixels in which Ca V 1.2 and EEA1 completely overlapped. b dot-plots summarizing % colocalization between EEA1 and Ca V 1.2 young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 3, n = 14; ISO: N = 3, n = 14) myocytes, and c EEA1-positive endosome areas in young (control: N = 3, n = 15; ISO: N = 3, n = 16) and old (control: N = 3, n = 14; ISO: N = 3, n = 13) myocytes. Data were analyzed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in ( b , c ) are from JAX young. Note no significant differences in EEA1/Ca V 1.2 colocalization, responsivity to ISO, or endosome size was detected when young JAX and young NIA myocytes were compared (Supplementary Fig. ). Data are presented as mean ± SEM. Source data are provided in the Source Data file.

    Techniques Used: Comparison

    a Representative whole-cell currents from shRNA-scrmb and shRNA-mBIN1 myocytes before and during ISO application. b fold change in peak I Ca with ISO in young, old, shRNA-scrmb ( N = 3, n = 7), and shRNA-mBIN1 ( N = 3, n = 9) myocytes. c Representative Ca 2+ transients recorded from old shRNA-scrmb and shRNA-mBIN1 myocytes before and after ISO.  Fold increase after ISO from young, old, shRNA-scrmb ( N = 3, n = 14), and shRNA-mBIN1 ( N = 5, n = 18) myocytes. e SMLM localization maps showing Ca V 1.2 channel localization on t-tubules of myocytes from old shRNA-scrmb and shRNA-mBIN1, with or without ISO stimulation. Regions of interest are highlighted by yellow boxes. f Fold change in mean Ca V 1.2 channel cluster area with ISO in the young, old, old shRNA-scrmb (control: N = 3, n = 14; ISO: N = 3, n = 13) and shRNA-mBIN1 myocytes (control: N = 3, n = 12; ISO: N = 3, n = 11). g , h show the same layout for RyR2 immunostained old shRNA-scrmb (control: N = 3, n = 12; ISO: N = 3, n = 13) and shRNA-mBIN1 myocytes (control: N = 3, n = 9; ISO: N = 3, n = 8). Old and young data points in ( b ,  , f , h ) are reproduced from data in Figs.  b, h, and  b,  respectively. Statistical analysis was performed on data in ( b ,  , f , h ) using one-way ANOVAs with multiple comparison post-hoc tests. Young data in ( b ) are pooled from NIA young and JAX young myocytes, data in (  , f , h ) are from NIA young myocytes. Note there was no significant difference in I Ca , Ca V 1.2, and RyR2 cluster areas when JAX and NIA young mice were compared (see Supplementary Fig.  ). Data are presented as mean ± SEM. Source data are provided in the Source Data file.
    Figure Legend Snippet: a Representative whole-cell currents from shRNA-scrmb and shRNA-mBIN1 myocytes before and during ISO application. b fold change in peak I Ca with ISO in young, old, shRNA-scrmb ( N = 3, n = 7), and shRNA-mBIN1 ( N = 3, n = 9) myocytes. c Representative Ca 2+ transients recorded from old shRNA-scrmb and shRNA-mBIN1 myocytes before and after ISO. Fold increase after ISO from young, old, shRNA-scrmb ( N = 3, n = 14), and shRNA-mBIN1 ( N = 5, n = 18) myocytes. e SMLM localization maps showing Ca V 1.2 channel localization on t-tubules of myocytes from old shRNA-scrmb and shRNA-mBIN1, with or without ISO stimulation. Regions of interest are highlighted by yellow boxes. f Fold change in mean Ca V 1.2 channel cluster area with ISO in the young, old, old shRNA-scrmb (control: N = 3, n = 14; ISO: N = 3, n = 13) and shRNA-mBIN1 myocytes (control: N = 3, n = 12; ISO: N = 3, n = 11). g , h show the same layout for RyR2 immunostained old shRNA-scrmb (control: N = 3, n = 12; ISO: N = 3, n = 13) and shRNA-mBIN1 myocytes (control: N = 3, n = 9; ISO: N = 3, n = 8). Old and young data points in ( b , , f , h ) are reproduced from data in Figs. b, h, and b, respectively. Statistical analysis was performed on data in ( b , , f , h ) using one-way ANOVAs with multiple comparison post-hoc tests. Young data in ( b ) are pooled from NIA young and JAX young myocytes, data in ( , f , h ) are from NIA young myocytes. Note there was no significant difference in I Ca , Ca V 1.2, and RyR2 cluster areas when JAX and NIA young mice were compared (see Supplementary Fig. ). Data are presented as mean ± SEM. Source data are provided in the Source Data file.

    Techniques Used: shRNA, Comparison

    The main findings of our study graphically illustrated and summarized. Top : In healthy young cells, Ca V 1.2 channels undergo endosomal recycling, where channels on endosomes are either marked for degradation through the late endosome pathway or are recycled to the sarcolemma through the fast and slow recycling pathways. Following β -adrenergic receptor ( β -AR) stimulation, a pool of channels localized to endosomes are mobilized to the membrane, resulting in larger Ca V 1.2 clusters along t-tubules. Across the dyad, RyR2 clusters on the sarcoplasmic reticulum also increase following βAR stimulation ensuring efficient Ca 2+ -induced Ca 2+ -release. This increase in cytosolic Ca 2+ , along with increased phosphorylation of cardiac Troponin I (cTnI) and cardiac myosin binding protein-C (cMyBP-C) within the sarcomere, allows for enhanced contractility under acute stress to cope with elevated hemodynamic and metabolic demands. Bottom : In aging, Bridging Integrator 1 (BIN1) protein levels are increased, accompanied by a swelling of endosomes and subsequent dysregulation of endosomal trafficking of Ca V 1.2. Ca V 1.2 and RyR2 channels are basally super-clustered at the dyads and lose β -AR responsivity. Reduced phosphorylation of cTnI and cMyBP-C result in systolic and diastolic dysfunction. BIN1 knockdown in aging recovers RyR2 clustering plasticity and Ca 2+ transient responsivity to β- AR stimulation. Phosphorylation of cMyBP-C is basally restored, and contractility is recovered to youthful levels. Thus, BIN1 knockdown rejuvenates the aging heart. Created with BioRender.com.
    Figure Legend Snippet: The main findings of our study graphically illustrated and summarized. Top : In healthy young cells, Ca V 1.2 channels undergo endosomal recycling, where channels on endosomes are either marked for degradation through the late endosome pathway or are recycled to the sarcolemma through the fast and slow recycling pathways. Following β -adrenergic receptor ( β -AR) stimulation, a pool of channels localized to endosomes are mobilized to the membrane, resulting in larger Ca V 1.2 clusters along t-tubules. Across the dyad, RyR2 clusters on the sarcoplasmic reticulum also increase following βAR stimulation ensuring efficient Ca 2+ -induced Ca 2+ -release. This increase in cytosolic Ca 2+ , along with increased phosphorylation of cardiac Troponin I (cTnI) and cardiac myosin binding protein-C (cMyBP-C) within the sarcomere, allows for enhanced contractility under acute stress to cope with elevated hemodynamic and metabolic demands. Bottom : In aging, Bridging Integrator 1 (BIN1) protein levels are increased, accompanied by a swelling of endosomes and subsequent dysregulation of endosomal trafficking of Ca V 1.2. Ca V 1.2 and RyR2 channels are basally super-clustered at the dyads and lose β -AR responsivity. Reduced phosphorylation of cTnI and cMyBP-C result in systolic and diastolic dysfunction. BIN1 knockdown in aging recovers RyR2 clustering plasticity and Ca 2+ transient responsivity to β- AR stimulation. Phosphorylation of cMyBP-C is basally restored, and contractility is recovered to youthful levels. Thus, BIN1 knockdown rejuvenates the aging heart. Created with BioRender.com.

    Techniques Used: Membrane, Binding Assay

    ca v 1 2  (Alomone Labs)


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    Alomone Labs ca v 1 2
    a . Western blot of hCO treated with ASO.14, ASO.17, ASO.18 or ASO.Scr (differentiation day 70-80). b . Ca V 1.2 protein blot normalized to GAPDH. Each dot represents an individual sample containing 2–3 hCO from an independent experiment (Ctrl: n = 8; TS Scr: n = 8; TS ASO.14: n = 6; TS ASO.17: n = 6; TS ASO.18: n = 6). Data are presented as mean ± s.d. One-way ANOVA with Tukey’s post hoc test: F 4,31 = 0.3548, P = 0.8387. c . Raw images of the western blots corresponding to a – b .
    Ca V 1 2, 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/ca v 1 2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ca v 1 2 - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "Antisense oligonucleotide therapeutic approach for Timothy syndrome"

    Article Title: Antisense oligonucleotide therapeutic approach for Timothy syndrome

    Journal: Nature

    doi: 10.1038/s41586-024-07310-6

    a . Western blot of hCO treated with ASO.14, ASO.17, ASO.18 or ASO.Scr (differentiation day 70-80). b . Ca V 1.2 protein blot normalized to GAPDH. Each dot represents an individual sample containing 2–3 hCO from an independent experiment (Ctrl: n = 8; TS Scr: n = 8; TS ASO.14: n = 6; TS ASO.17: n = 6; TS ASO.18: n = 6). Data are presented as mean ± s.d. One-way ANOVA with Tukey’s post hoc test: F 4,31 = 0.3548, P = 0.8387. c . Raw images of the western blots corresponding to a – b .
    Figure Legend Snippet: a . Western blot of hCO treated with ASO.14, ASO.17, ASO.18 or ASO.Scr (differentiation day 70-80). b . Ca V 1.2 protein blot normalized to GAPDH. Each dot represents an individual sample containing 2–3 hCO from an independent experiment (Ctrl: n = 8; TS Scr: n = 8; TS ASO.14: n = 6; TS ASO.17: n = 6; TS ASO.18: n = 6). Data are presented as mean ± s.d. One-way ANOVA with Tukey’s post hoc test: F 4,31 = 0.3548, P = 0.8387. c . Raw images of the western blots corresponding to a – b .

    Techniques Used: Western Blot

    a . Experimental design. Each condition contains a mix of two plasmids encoding either the WT or the TS Ca V 1.2 that are co-transfected with plasmids encoding the β1b, α2δ subunit of the channel plus GCaMP6-x. b . Representative images of GCaMP imaging before and after 67 mM KCl application. Transfected cells contain only WT Ca V 1.2 (upper panel) or only TS Ca V 1.2 (lower panel). Scale bar 100 μm. c . Representative traces of chemically induced intracellular GCaMP signal. (0% TS + 100% WT, n = 257 cells; 2.5% TS + 97.5% WT, n = 388 cells; 7.5% TS + 92.5% WT, n = 335 cells; 17.5% TS + 82.5% WT, n = 325 cells; 100% TS + 0% WT, n = 190 cells). Data are presented as mean ± s.e.m. d . Comparison of residual Ca 2+ measured after 67 mM KCl application. Each dot represents one cell (n = 11,228 cells). One-way ANOVA with Dunnett’s correction was used was used to compare TS Ca V 1.2 to WT: F 11, 11266 = 68.73, P < 0.0001, ****P < 0.0001. e . Experimental procedure for evaluating rescue by ASOs at various concentrations. f . Representative images of GCaMP6f imaging acquired before and after 67 mM KCl application to control (upper panel) and TS neurons (lower panel) neurons. Control and TS neurons are derived from isogenic hiPS cells (242 to 254 days of differentiation). Scale bar 50 μm. g . Comparison of residual Ca 2+ measured by GCaMP6f after 67 mM KCl exposure. Each dot represents one cell (n = 1,527 cells). One-way ANOVA with Dunnett’s correction was used to compare ASO.14, ASO.17 and ASO.18 to TS ASO.Scr: F 13, 1513 = 20.45, P < 0.0001; ****P < 0.0001.
    Figure Legend Snippet: a . Experimental design. Each condition contains a mix of two plasmids encoding either the WT or the TS Ca V 1.2 that are co-transfected with plasmids encoding the β1b, α2δ subunit of the channel plus GCaMP6-x. b . Representative images of GCaMP imaging before and after 67 mM KCl application. Transfected cells contain only WT Ca V 1.2 (upper panel) or only TS Ca V 1.2 (lower panel). Scale bar 100 μm. c . Representative traces of chemically induced intracellular GCaMP signal. (0% TS + 100% WT, n = 257 cells; 2.5% TS + 97.5% WT, n = 388 cells; 7.5% TS + 92.5% WT, n = 335 cells; 17.5% TS + 82.5% WT, n = 325 cells; 100% TS + 0% WT, n = 190 cells). Data are presented as mean ± s.e.m. d . Comparison of residual Ca 2+ measured after 67 mM KCl application. Each dot represents one cell (n = 11,228 cells). One-way ANOVA with Dunnett’s correction was used was used to compare TS Ca V 1.2 to WT: F 11, 11266 = 68.73, P < 0.0001, ****P < 0.0001. e . Experimental procedure for evaluating rescue by ASOs at various concentrations. f . Representative images of GCaMP6f imaging acquired before and after 67 mM KCl application to control (upper panel) and TS neurons (lower panel) neurons. Control and TS neurons are derived from isogenic hiPS cells (242 to 254 days of differentiation). Scale bar 50 μm. g . Comparison of residual Ca 2+ measured by GCaMP6f after 67 mM KCl exposure. Each dot represents one cell (n = 1,527 cells). One-way ANOVA with Dunnett’s correction was used to compare ASO.14, ASO.17 and ASO.18 to TS ASO.Scr: F 13, 1513 = 20.45, P < 0.0001; ****P < 0.0001.

    Techniques Used: Transfection, Imaging, Comparison, Derivative Assay

    ca v 1 2  (Alomone Labs)


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    Structured Review

    Alomone Labs ca v 1 2
    The cascade is initiated by the binding of an agonist (e.g. epinephrine or norepinephrine; depicted as a small green circle) to a β-adrenergic receptor (βAR) which, in turn, activates the G protein G s by catalyzing GDP-GTP exchange at the Gα s subunit followed by separation of the latter from the Gβγ subunit. The GTP-bound Gα s , alone or in concert with Gβγ, activates adenylyl cyclase (AC), promoting the conversion of ATP to cAMP. cAMP activates PKA, causing the dissociation of its regulatory (RS) from catalytic (CS) subunits. CS phosphorylates several targets including α 1C and Ca V β subunits, but the most important target in Ca V 1.2 regulation appears to be the Rad protein. Phosphorylation of Rad removes the constitutive inhibition that it exerts upon channel activity, through separation of Rad from Ca V β. Before activation, the PKA holoenzyme is thought to be associated with the α 1C subunit via an AKAP protein that strongly binds PKAR (the binding main sites for most AKAPs are in the dCT of α 1C ). The scheme also shows the auxiliary subunits of Ca V 1.2 and the cytosolic domain of α 1C : N-terminus (NT), C-terminus (CT) (with dCT colored dark mustard), and intracellular loops (shown in light gray). Also indicated are the approximate CT proteolytic cleavage site (scissors), major PKA phosphorylation sites in the CT (S1700 and S1928), and the DCRD, CBS, and PCRD domains.
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    1) Product Images from "Tripartite interactions of PKA catalytic subunit and C-terminal domains of cardiac Ca 2+ channel modulate its β-adrenergic regulation"

    Article Title: Tripartite interactions of PKA catalytic subunit and C-terminal domains of cardiac Ca 2+ channel modulate its β-adrenergic regulation

    Journal: bioRxiv

    doi: 10.1101/2023.11.28.564875

    The cascade is initiated by the binding of an agonist (e.g. epinephrine or norepinephrine; depicted as a small green circle) to a β-adrenergic receptor (βAR) which, in turn, activates the G protein G s by catalyzing GDP-GTP exchange at the Gα s subunit followed by separation of the latter from the Gβγ subunit. The GTP-bound Gα s , alone or in concert with Gβγ, activates adenylyl cyclase (AC), promoting the conversion of ATP to cAMP. cAMP activates PKA, causing the dissociation of its regulatory (RS) from catalytic (CS) subunits. CS phosphorylates several targets including α 1C and Ca V β subunits, but the most important target in Ca V 1.2 regulation appears to be the Rad protein. Phosphorylation of Rad removes the constitutive inhibition that it exerts upon channel activity, through separation of Rad from Ca V β. Before activation, the PKA holoenzyme is thought to be associated with the α 1C subunit via an AKAP protein that strongly binds PKAR (the binding main sites for most AKAPs are in the dCT of α 1C ). The scheme also shows the auxiliary subunits of Ca V 1.2 and the cytosolic domain of α 1C : N-terminus (NT), C-terminus (CT) (with dCT colored dark mustard), and intracellular loops (shown in light gray). Also indicated are the approximate CT proteolytic cleavage site (scissors), major PKA phosphorylation sites in the CT (S1700 and S1928), and the DCRD, CBS, and PCRD domains.
    Figure Legend Snippet: The cascade is initiated by the binding of an agonist (e.g. epinephrine or norepinephrine; depicted as a small green circle) to a β-adrenergic receptor (βAR) which, in turn, activates the G protein G s by catalyzing GDP-GTP exchange at the Gα s subunit followed by separation of the latter from the Gβγ subunit. The GTP-bound Gα s , alone or in concert with Gβγ, activates adenylyl cyclase (AC), promoting the conversion of ATP to cAMP. cAMP activates PKA, causing the dissociation of its regulatory (RS) from catalytic (CS) subunits. CS phosphorylates several targets including α 1C and Ca V β subunits, but the most important target in Ca V 1.2 regulation appears to be the Rad protein. Phosphorylation of Rad removes the constitutive inhibition that it exerts upon channel activity, through separation of Rad from Ca V β. Before activation, the PKA holoenzyme is thought to be associated with the α 1C subunit via an AKAP protein that strongly binds PKAR (the binding main sites for most AKAPs are in the dCT of α 1C ). The scheme also shows the auxiliary subunits of Ca V 1.2 and the cytosolic domain of α 1C : N-terminus (NT), C-terminus (CT) (with dCT colored dark mustard), and intracellular loops (shown in light gray). Also indicated are the approximate CT proteolytic cleavage site (scissors), major PKA phosphorylation sites in the CT (S1700 and S1928), and the DCRD, CBS, and PCRD domains.

    Techniques Used: Binding Assay, Inhibition, Activity Assay, Activation Assay

    The original data from Katz et al. paper  were analyzed in the same way as shown in Fig. 8, to seek for correlation between I Ba and the extent of current increase caused by Iso (50 µM Iso were used in this work). Oocytes expressed all Ca V 1.2 subunits (α 1C , β2b, α2δ), rad and β1AR. A and B , correlation between basal I Ba and fold increase by Iso in individual cells. Parameters of Spearman correlation analysis for the two distributions are shown in insets. Note that the amplitudes of I Ba in Ca V 1.2 with FL-α 1C were smaller (<0.7 µA) than for α 1C Δ1821 (0.1-2 µA). C , Comparison of the differences in Iso-induced increase in I Ba of α 1C Δ1821 and α 1C -FL for basal I Ba of small-intermediate and high amplitudes. In this series of experiments large (>0.7 µA) currents were observed only with α 1C Δ1821. Despite the smaller number of measurements, the tendency observed in experiments of Fig. 8E is also observed here: the Iso-induced increase in I Ba is larger for α 1C Δ1821 than in α 1C -FL for smaller currents, but not for larger α 1C Δ1821 currents.
    Figure Legend Snippet: The original data from Katz et al. paper were analyzed in the same way as shown in Fig. 8, to seek for correlation between I Ba and the extent of current increase caused by Iso (50 µM Iso were used in this work). Oocytes expressed all Ca V 1.2 subunits (α 1C , β2b, α2δ), rad and β1AR. A and B , correlation between basal I Ba and fold increase by Iso in individual cells. Parameters of Spearman correlation analysis for the two distributions are shown in insets. Note that the amplitudes of I Ba in Ca V 1.2 with FL-α 1C were smaller (<0.7 µA) than for α 1C Δ1821 (0.1-2 µA). C , Comparison of the differences in Iso-induced increase in I Ba of α 1C Δ1821 and α 1C -FL for basal I Ba of small-intermediate and high amplitudes. In this series of experiments large (>0.7 µA) currents were observed only with α 1C Δ1821. Despite the smaller number of measurements, the tendency observed in experiments of Fig. 8E is also observed here: the Iso-induced increase in I Ba is larger for α 1C Δ1821 than in α 1C -FL for smaller currents, but not for larger α 1C Δ1821 currents.

    Techniques Used: Comparison

    anti ca v 1 2 antibody  (Alomone Labs)


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    Alomone Labs anti ca v 1 2 antibody
    The protection of Ahf-caltide on the Langerdoff-perfused rat heart subjected to H 2 O 2 treatment. ( A ) Schematic diagram of the experimental protocol. ( B , C ) Time course of changes in heart rate ( B ) and coronary flow ( C ) among groups. ( D ) LDH activity of coronary effluent in groups. ( E , F ) Representatives immunoblots ( E ) and quantitative analysis ( F ) of Ca V 1.2, Ca V β2, CAST and BNP levels in hearts of groups. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. control group; # p < 0.05 and ## p < 0.01 vs. H 2 O 2 group. NS, not significant. Results represent at least five independent experiments.
    Anti Ca V 1 2 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
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    1) Product Images from "Ahf-Caltide, a Novel Polypeptide Derived from Calpastatin, Protects against Oxidative Stress Injury by Stabilizing the Expression of Ca V 1.2 Calcium Channel"

    Article Title: Ahf-Caltide, a Novel Polypeptide Derived from Calpastatin, Protects against Oxidative Stress Injury by Stabilizing the Expression of Ca V 1.2 Calcium Channel

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms242115729

    The protection of Ahf-caltide on the Langerdoff-perfused rat heart subjected to H 2 O 2 treatment. ( A ) Schematic diagram of the experimental protocol. ( B , C ) Time course of changes in heart rate ( B ) and coronary flow ( C ) among groups. ( D ) LDH activity of coronary effluent in groups. ( E , F ) Representatives immunoblots ( E ) and quantitative analysis ( F ) of Ca V 1.2, Ca V β2, CAST and BNP levels in hearts of groups. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. control group; # p < 0.05 and ## p < 0.01 vs. H 2 O 2 group. NS, not significant. Results represent at least five independent experiments.
    Figure Legend Snippet: The protection of Ahf-caltide on the Langerdoff-perfused rat heart subjected to H 2 O 2 treatment. ( A ) Schematic diagram of the experimental protocol. ( B , C ) Time course of changes in heart rate ( B ) and coronary flow ( C ) among groups. ( D ) LDH activity of coronary effluent in groups. ( E , F ) Representatives immunoblots ( E ) and quantitative analysis ( F ) of Ca V 1.2, Ca V β2, CAST and BNP levels in hearts of groups. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. control group; # p < 0.05 and ## p < 0.01 vs. H 2 O 2 group. NS, not significant. Results represent at least five independent experiments.

    Techniques Used: Activity Assay, Western Blot

    The protection of Ahf-caltide on the Langerdoff-perfused rat hearts subjected to IR injury. ( A ) Schematic diagram of the experimental protocol. ( B , C ) Time course of changes in heart rate ( B ) and coronary flow ( C ) among groups. ( D ) LDH activity of coronary effluent in groups. ( E , F ) The SOD activity ( E ) and MDA content ( F ) in the heart tissues. ( G , H ) Representative images ( G ) and quantitative analysis ( H ) of Ca V 1.2, Ca V β2 and CAST in hearts among groups. * p < 0.05 and *** p < 0.001 vs. control group; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. IR group. NS, not significant. Results represent at least five independent experiments.
    Figure Legend Snippet: The protection of Ahf-caltide on the Langerdoff-perfused rat hearts subjected to IR injury. ( A ) Schematic diagram of the experimental protocol. ( B , C ) Time course of changes in heart rate ( B ) and coronary flow ( C ) among groups. ( D ) LDH activity of coronary effluent in groups. ( E , F ) The SOD activity ( E ) and MDA content ( F ) in the heart tissues. ( G , H ) Representative images ( G ) and quantitative analysis ( H ) of Ca V 1.2, Ca V β2 and CAST in hearts among groups. * p < 0.05 and *** p < 0.001 vs. control group; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. IR group. NS, not significant. Results represent at least five independent experiments.

    Techniques Used: Activity Assay

    The protection by Ahf-caltide on H9c2 cell injury induced by CoCl 2 treatment. ( A ) The localization of Ahf-caltide in H9c2 cells examined by fluorescence microscopy (scale bars, 100 μm). ( B ) The cell viability was detected by CCK-8. ( C ) LDH activity of supernatant in groups. ( D , E ) ROS level in groups (scale bars, 100 μm). ( F , G ) Representative immunoblots ( F ) and quantitative analysis ( G ) of expression levels of Ca V 1.2, Ca V β2, CAST in groups. ( H , I ) Representative immunoblots ( H ) and quantitative analysis ( I ) of the effects of cycloheximide (CHX) on the expression of Ca V 1.2, Ca V β2, CAST in groups. ( J , K ) Representative immunoblots ( J ) and quantitative analysis ( K ) of the effects of MG-132/Leupeptin on the levels of Ca V 1.2, Ca V β2, CAST of groups. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. control group or as indicated on the figure; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. CoCl 2 group. NS, no significance. Results represent at least three independent experiments.
    Figure Legend Snippet: The protection by Ahf-caltide on H9c2 cell injury induced by CoCl 2 treatment. ( A ) The localization of Ahf-caltide in H9c2 cells examined by fluorescence microscopy (scale bars, 100 μm). ( B ) The cell viability was detected by CCK-8. ( C ) LDH activity of supernatant in groups. ( D , E ) ROS level in groups (scale bars, 100 μm). ( F , G ) Representative immunoblots ( F ) and quantitative analysis ( G ) of expression levels of Ca V 1.2, Ca V β2, CAST in groups. ( H , I ) Representative immunoblots ( H ) and quantitative analysis ( I ) of the effects of cycloheximide (CHX) on the expression of Ca V 1.2, Ca V β2, CAST in groups. ( J , K ) Representative immunoblots ( J ) and quantitative analysis ( K ) of the effects of MG-132/Leupeptin on the levels of Ca V 1.2, Ca V β2, CAST of groups. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. control group or as indicated on the figure; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. CoCl 2 group. NS, no significance. Results represent at least three independent experiments.

    Techniques Used: Fluorescence, Microscopy, CCK-8 Assay, Activity Assay, Western Blot, Expressing

    Concentration-dependent effect of H 2 O 2 on CS L binding to N- and C-terminus of Ca V 1.2 channel α 1 subunit. GST pull-down assay for the effect of H 2 O 2 (10 −1 –10 −6 M) on the binding of CS L to CT1 ( A ) or PreIQ ( C ) or IQ ( E ) or NT ( K ). Plots of the effect of H 2 O 2 with fitted curves on the CS L binding to CT1 ( B ) or PreIQ ( D ) or IQ ( F ) or NT ( L ). Pull-down assay ( G ) and binding curves ( H ) for CS L binding to NT. Pull-down assay for the effect of [Ca 2+ ] ( I ) and quantitative analysis ( J ) on the binding of CS L to NT. Symbols are as indicated in each graph. NS, no significance. Protein bands were visualized using Coomassie brilliant blue staining and indicated by arrows. Results represent at least four independent experiments.
    Figure Legend Snippet: Concentration-dependent effect of H 2 O 2 on CS L binding to N- and C-terminus of Ca V 1.2 channel α 1 subunit. GST pull-down assay for the effect of H 2 O 2 (10 −1 –10 −6 M) on the binding of CS L to CT1 ( A ) or PreIQ ( C ) or IQ ( E ) or NT ( K ). Plots of the effect of H 2 O 2 with fitted curves on the CS L binding to CT1 ( B ) or PreIQ ( D ) or IQ ( F ) or NT ( L ). Pull-down assay ( G ) and binding curves ( H ) for CS L binding to NT. Pull-down assay for the effect of [Ca 2+ ] ( I ) and quantitative analysis ( J ) on the binding of CS L to NT. Symbols are as indicated in each graph. NS, no significance. Protein bands were visualized using Coomassie brilliant blue staining and indicated by arrows. Results represent at least four independent experiments.

    Techniques Used: Concentration Assay, Binding Assay, Pull Down Assay, Staining

    Molecular docking analysis of CS L or CS L 54-64 binding to N- and C- terminus of Ca V 1.2 channel α 1 subunit. ( A – C ) Improved five groups of epitope clusters in  corresponding to the totally docked poses of ( A1 ) CS L /PreIQ, ( B1 ) CS L /IQ, ( C1 ) CS L /NT; the fingerprints for the interaction obtained from the five groups’ docked poses between CS L and ( A2 ) PreIQ, ( B2 ) IQ, ( C2 ) NT; the fingerprints for the interaction from the first group pose between CS L and ( A3 ) PreIQ, ( B3 ) IQ, ( C3 ) NT; the docked poses of CS L 54-64 binding to ( A4 ) PreIQ, ( B4 ) IQ, ( C4 ) NT; the interaction diagram of Lys 64, Lys 56 and Glu 54 on CS L 54-64 with PreIQ ( A5 ), IQ ( B5 ) and NT ( C5 ), respectively. The diagrams in the first group with the highest score are represented by blue “spaghetti” and the others with a translucent “spaghetti”. Blue structures represent PreIQ/IQ/NT, red structures represent CS L or CS L 54-64.
    Figure Legend Snippet: Molecular docking analysis of CS L or CS L 54-64 binding to N- and C- terminus of Ca V 1.2 channel α 1 subunit. ( A – C ) Improved five groups of epitope clusters in corresponding to the totally docked poses of ( A1 ) CS L /PreIQ, ( B1 ) CS L /IQ, ( C1 ) CS L /NT; the fingerprints for the interaction obtained from the five groups’ docked poses between CS L and ( A2 ) PreIQ, ( B2 ) IQ, ( C2 ) NT; the fingerprints for the interaction from the first group pose between CS L and ( A3 ) PreIQ, ( B3 ) IQ, ( C3 ) NT; the docked poses of CS L 54-64 binding to ( A4 ) PreIQ, ( B4 ) IQ, ( C4 ) NT; the interaction diagram of Lys 64, Lys 56 and Glu 54 on CS L 54-64 with PreIQ ( A5 ), IQ ( B5 ) and NT ( C5 ), respectively. The diagrams in the first group with the highest score are represented by blue “spaghetti” and the others with a translucent “spaghetti”. Blue structures represent PreIQ/IQ/NT, red structures represent CS L or CS L 54-64.

    Techniques Used: Binding Assay


    Figure Legend Snippet: Predicted binding sites of CS L 54-65 to Ca V 1.2 channels.

    Techniques Used: Binding Assay

    anti ca v 3 2 rabbit  (Alomone Labs)


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    Alomone Labs anti ca v 3 2 rabbit
    D + Q turnover caveolar Ca 2+ sparks in aged mesenteric VSMCs. (a), experimental design of senolytics administration is shown. (D, dasatinib; Q, quercetin; PEG, polyethylene glycol). (b), heat map of RNA‐seq data of mesenteric artery from young, vehicle‐, and D + Q‐treatment mice ( n = 4 samples for each group). (c–f) Shown are gene set enrichment analyses (c, d) and heat maps for the top 20 up‐regulated genes for CELL CYCLE (e) and APOPTOSIS (f) in D + Q compared with vehicle group. FDR, false discovery rate. (g), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle‐aged mouse and the time course of Ca 2+ fluorescence changes. (h), same as (g) but in the presence of methyl‐β‐cyclodextrin (dextrin, 10 mM, 90 min at room temperature). (i), same as (g) but in the cell from a D + Q‐treated aged mouse. (j), same as (i) but in the presence of methyl‐β‐cyclodextrin. (k–l), summary of the results. Ca 2+ spark frequency (e) and fraction of cells producing Ca 2+ sparks (f) in VSMCs from aged mice ( n = 134), in VSMCs from aged mice incubated with methyl‐β‐cyclodextrin ( n = 95), in VSMCs from D + Q treated aged mice ( n = 175), and in VSMCs from D + Q treated aged mice incubated with methyl‐β‐cyclodextrin ( n = 175). Cells were isolated from 4 mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05 ; n.s., not significant. (m), western blot analysis of Ca V 3.2, Caveolin‐1 proteins in mesenteric arteries of aged versus D + Q mice. (n), quantification of western blot results. Western blot results were analyzed from 8 mice in each group. * p < 0.05 ; n.s., not significant; Cav‐1, Caveolin‐1.
    Anti Ca V 3 2 Rabbit, 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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    1) Product Images from "Senolytics prevent caveolar Ca V 3 . 2‐RyR axis malfunction in old vascular smooth muscle"

    Article Title: Senolytics prevent caveolar Ca V 3 . 2‐RyR axis malfunction in old vascular smooth muscle

    Journal: Aging Cell

    doi: 10.1111/acel.14002

    D + Q turnover caveolar Ca 2+ sparks in aged mesenteric VSMCs. (a), experimental design of senolytics administration is shown. (D, dasatinib; Q, quercetin; PEG, polyethylene glycol). (b), heat map of RNA‐seq data of mesenteric artery from young, vehicle‐, and D + Q‐treatment mice ( n = 4 samples for each group). (c–f) Shown are gene set enrichment analyses (c, d) and heat maps for the top 20 up‐regulated genes for CELL CYCLE (e) and APOPTOSIS (f) in D + Q compared with vehicle group. FDR, false discovery rate. (g), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle‐aged mouse and the time course of Ca 2+ fluorescence changes. (h), same as (g) but in the presence of methyl‐β‐cyclodextrin (dextrin, 10 mM, 90 min at room temperature). (i), same as (g) but in the cell from a D + Q‐treated aged mouse. (j), same as (i) but in the presence of methyl‐β‐cyclodextrin. (k–l), summary of the results. Ca 2+ spark frequency (e) and fraction of cells producing Ca 2+ sparks (f) in VSMCs from aged mice ( n = 134), in VSMCs from aged mice incubated with methyl‐β‐cyclodextrin ( n = 95), in VSMCs from D + Q treated aged mice ( n = 175), and in VSMCs from D + Q treated aged mice incubated with methyl‐β‐cyclodextrin ( n = 175). Cells were isolated from 4 mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05 ; n.s., not significant. (m), western blot analysis of Ca V 3.2, Caveolin‐1 proteins in mesenteric arteries of aged versus D + Q mice. (n), quantification of western blot results. Western blot results were analyzed from 8 mice in each group. * p < 0.05 ; n.s., not significant; Cav‐1, Caveolin‐1.
    Figure Legend Snippet: D + Q turnover caveolar Ca 2+ sparks in aged mesenteric VSMCs. (a), experimental design of senolytics administration is shown. (D, dasatinib; Q, quercetin; PEG, polyethylene glycol). (b), heat map of RNA‐seq data of mesenteric artery from young, vehicle‐, and D + Q‐treatment mice ( n = 4 samples for each group). (c–f) Shown are gene set enrichment analyses (c, d) and heat maps for the top 20 up‐regulated genes for CELL CYCLE (e) and APOPTOSIS (f) in D + Q compared with vehicle group. FDR, false discovery rate. (g), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle‐aged mouse and the time course of Ca 2+ fluorescence changes. (h), same as (g) but in the presence of methyl‐β‐cyclodextrin (dextrin, 10 mM, 90 min at room temperature). (i), same as (g) but in the cell from a D + Q‐treated aged mouse. (j), same as (i) but in the presence of methyl‐β‐cyclodextrin. (k–l), summary of the results. Ca 2+ spark frequency (e) and fraction of cells producing Ca 2+ sparks (f) in VSMCs from aged mice ( n = 134), in VSMCs from aged mice incubated with methyl‐β‐cyclodextrin ( n = 95), in VSMCs from D + Q treated aged mice ( n = 175), and in VSMCs from D + Q treated aged mice incubated with methyl‐β‐cyclodextrin ( n = 175). Cells were isolated from 4 mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05 ; n.s., not significant. (m), western blot analysis of Ca V 3.2, Caveolin‐1 proteins in mesenteric arteries of aged versus D + Q mice. (n), quantification of western blot results. Western blot results were analyzed from 8 mice in each group. * p < 0.05 ; n.s., not significant; Cav‐1, Caveolin‐1.

    Techniques Used: RNA Sequencing Assay, Fluorescence, Incubation, Isolation, Western Blot

    D + Q promote caveolae remodeling and Ca V 3.2‐Cav‐1 co‐localization to rescue caveolar Ca V 3.2‐RyR axis in aged VSMC. (a–c), shown are gene set enrichment analyses (a) and heat maps for the top 10 up‐regulated genes (b, c) for ATP BIOSYNTHETIC PROCESS and MICROTUBULE CYTOSKELETON ORGANIZATION in D + Q compared with vehicle group. (d, e), electron microscopy image of a vehicle and a D + Q treated VSMC, and summary of the results. Caveolae density, diameter of caveolae neck in VSMCs from vehicle‐ versus D + Q‐treated mice (four mice in each group). (f‐g), confocal immunofluorescence images and the line course of fluorescence changes. Immunofluorescently labeled with Ca V 3.2 (green) and Cav‐1 (red) in mesenteric arteries from vehicle‐ and D + Q‐treated mice. Bar, 20 μm. (h), Pearson's correlation coefficients for colocalization assays. The plot shows Pearson's correlation coefficients for the colocalization analysis ( n = 30–40 arteries). The Kruskal–Wallis H test was used for calculating statistical differences. Arteries were isolated from four mice in each group. (i), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle aged mouse and the time course of Ca 2+ fluorescence changes. (j), same as (h) but in the presence of Ni 2+ (50 μM). (k), same as (h) but in the cell from a D + Q treated aged mouse. (l), same as (j) but in the presence of Ni 2+ . (m–p), summary of the results. Ca 2+ spark frequency (m) and fraction of cells producing Ca 2+ sparks (n) in VSMCs from aged mice ( n = 124), in VSMCs from aged mice incubated with Ni 2+ ( n = 109), in VSMCs from D + Q‐treated aged mice ( n = 176), and in VSMCs from D + Q‐treated aged mice incubated with Ni 2+ ( n = 115). Ca 2+ spark frequency (o) and fraction of cells producing Ca 2+ sparks (p) in VSMCs from D + Q treated aged mice incubated with Ni 2+ ( n = 112), with methyl‐β‐cyclodextrin ( n = 108), as well as Ni 2+ +methyl‐β‐cyclodextrin ( n = 96). Cells were isolated from four mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05; n.s., not significant.
    Figure Legend Snippet: D + Q promote caveolae remodeling and Ca V 3.2‐Cav‐1 co‐localization to rescue caveolar Ca V 3.2‐RyR axis in aged VSMC. (a–c), shown are gene set enrichment analyses (a) and heat maps for the top 10 up‐regulated genes (b, c) for ATP BIOSYNTHETIC PROCESS and MICROTUBULE CYTOSKELETON ORGANIZATION in D + Q compared with vehicle group. (d, e), electron microscopy image of a vehicle and a D + Q treated VSMC, and summary of the results. Caveolae density, diameter of caveolae neck in VSMCs from vehicle‐ versus D + Q‐treated mice (four mice in each group). (f‐g), confocal immunofluorescence images and the line course of fluorescence changes. Immunofluorescently labeled with Ca V 3.2 (green) and Cav‐1 (red) in mesenteric arteries from vehicle‐ and D + Q‐treated mice. Bar, 20 μm. (h), Pearson's correlation coefficients for colocalization assays. The plot shows Pearson's correlation coefficients for the colocalization analysis ( n = 30–40 arteries). The Kruskal–Wallis H test was used for calculating statistical differences. Arteries were isolated from four mice in each group. (i), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle aged mouse and the time course of Ca 2+ fluorescence changes. (j), same as (h) but in the presence of Ni 2+ (50 μM). (k), same as (h) but in the cell from a D + Q treated aged mouse. (l), same as (j) but in the presence of Ni 2+ . (m–p), summary of the results. Ca 2+ spark frequency (m) and fraction of cells producing Ca 2+ sparks (n) in VSMCs from aged mice ( n = 124), in VSMCs from aged mice incubated with Ni 2+ ( n = 109), in VSMCs from D + Q‐treated aged mice ( n = 176), and in VSMCs from D + Q‐treated aged mice incubated with Ni 2+ ( n = 115). Ca 2+ spark frequency (o) and fraction of cells producing Ca 2+ sparks (p) in VSMCs from D + Q treated aged mice incubated with Ni 2+ ( n = 112), with methyl‐β‐cyclodextrin ( n = 108), as well as Ni 2+ +methyl‐β‐cyclodextrin ( n = 96). Cells were isolated from four mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05; n.s., not significant.

    Techniques Used: Electron Microscopy, Immunofluorescence, Fluorescence, Labeling, Isolation, Incubation

    anti ca v 3 2 rabbit  (Alomone Labs)


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    Alomone Labs anti ca v 3 2 rabbit
    D + Q turnover caveolar Ca 2+ sparks in aged mesenteric VSMCs. (a), experimental design of senolytics administration is shown. (D, dasatinib; Q, quercetin; PEG, polyethylene glycol). (b), heat map of RNA‐seq data of mesenteric artery from young, vehicle‐, and D + Q‐treatment mice ( n = 4 samples for each group). (c–f) Shown are gene set enrichment analyses (c, d) and heat maps for the top 20 up‐regulated genes for CELL CYCLE (e) and APOPTOSIS (f) in D + Q compared with vehicle group. FDR, false discovery rate. (g), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle‐aged mouse and the time course of Ca 2+ fluorescence changes. (h), same as (g) but in the presence of methyl‐β‐cyclodextrin (dextrin, 10 mM, 90 min at room temperature). (i), same as (g) but in the cell from a D + Q‐treated aged mouse. (j), same as (i) but in the presence of methyl‐β‐cyclodextrin. (k–l), summary of the results. Ca 2+ spark frequency (e) and fraction of cells producing Ca 2+ sparks (f) in VSMCs from aged mice ( n = 134), in VSMCs from aged mice incubated with methyl‐β‐cyclodextrin ( n = 95), in VSMCs from D + Q treated aged mice ( n = 175), and in VSMCs from D + Q treated aged mice incubated with methyl‐β‐cyclodextrin ( n = 175). Cells were isolated from 4 mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05 ; n.s., not significant. (m), western blot analysis of Ca V 3.2, Caveolin‐1 proteins in mesenteric arteries of aged versus D + Q mice. (n), quantification of western blot results. Western blot results were analyzed from 8 mice in each group. * p < 0.05 ; n.s., not significant; Cav‐1, Caveolin‐1.
    Anti Ca V 3 2 Rabbit, 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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    1) Product Images from "Senolytics prevent caveolar Ca V 3 . 2‐RyR axis malfunction in old vascular smooth muscle"

    Article Title: Senolytics prevent caveolar Ca V 3 . 2‐RyR axis malfunction in old vascular smooth muscle

    Journal: Aging Cell

    doi: 10.1111/acel.14002

    D + Q turnover caveolar Ca 2+ sparks in aged mesenteric VSMCs. (a), experimental design of senolytics administration is shown. (D, dasatinib; Q, quercetin; PEG, polyethylene glycol). (b), heat map of RNA‐seq data of mesenteric artery from young, vehicle‐, and D + Q‐treatment mice ( n = 4 samples for each group). (c–f) Shown are gene set enrichment analyses (c, d) and heat maps for the top 20 up‐regulated genes for CELL CYCLE (e) and APOPTOSIS (f) in D + Q compared with vehicle group. FDR, false discovery rate. (g), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle‐aged mouse and the time course of Ca 2+ fluorescence changes. (h), same as (g) but in the presence of methyl‐β‐cyclodextrin (dextrin, 10 mM, 90 min at room temperature). (i), same as (g) but in the cell from a D + Q‐treated aged mouse. (j), same as (i) but in the presence of methyl‐β‐cyclodextrin. (k–l), summary of the results. Ca 2+ spark frequency (e) and fraction of cells producing Ca 2+ sparks (f) in VSMCs from aged mice ( n = 134), in VSMCs from aged mice incubated with methyl‐β‐cyclodextrin ( n = 95), in VSMCs from D + Q treated aged mice ( n = 175), and in VSMCs from D + Q treated aged mice incubated with methyl‐β‐cyclodextrin ( n = 175). Cells were isolated from 4 mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05 ; n.s., not significant. (m), western blot analysis of Ca V 3.2, Caveolin‐1 proteins in mesenteric arteries of aged versus D + Q mice. (n), quantification of western blot results. Western blot results were analyzed from 8 mice in each group. * p < 0.05 ; n.s., not significant; Cav‐1, Caveolin‐1.
    Figure Legend Snippet: D + Q turnover caveolar Ca 2+ sparks in aged mesenteric VSMCs. (a), experimental design of senolytics administration is shown. (D, dasatinib; Q, quercetin; PEG, polyethylene glycol). (b), heat map of RNA‐seq data of mesenteric artery from young, vehicle‐, and D + Q‐treatment mice ( n = 4 samples for each group). (c–f) Shown are gene set enrichment analyses (c, d) and heat maps for the top 20 up‐regulated genes for CELL CYCLE (e) and APOPTOSIS (f) in D + Q compared with vehicle group. FDR, false discovery rate. (g), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle‐aged mouse and the time course of Ca 2+ fluorescence changes. (h), same as (g) but in the presence of methyl‐β‐cyclodextrin (dextrin, 10 mM, 90 min at room temperature). (i), same as (g) but in the cell from a D + Q‐treated aged mouse. (j), same as (i) but in the presence of methyl‐β‐cyclodextrin. (k–l), summary of the results. Ca 2+ spark frequency (e) and fraction of cells producing Ca 2+ sparks (f) in VSMCs from aged mice ( n = 134), in VSMCs from aged mice incubated with methyl‐β‐cyclodextrin ( n = 95), in VSMCs from D + Q treated aged mice ( n = 175), and in VSMCs from D + Q treated aged mice incubated with methyl‐β‐cyclodextrin ( n = 175). Cells were isolated from 4 mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05 ; n.s., not significant. (m), western blot analysis of Ca V 3.2, Caveolin‐1 proteins in mesenteric arteries of aged versus D + Q mice. (n), quantification of western blot results. Western blot results were analyzed from 8 mice in each group. * p < 0.05 ; n.s., not significant; Cav‐1, Caveolin‐1.

    Techniques Used: RNA Sequencing Assay, Fluorescence, Incubation, Isolation, Western Blot

    D + Q promote caveolae remodeling and Ca V 3.2‐Cav‐1 co‐localization to rescue caveolar Ca V 3.2‐RyR axis in aged VSMC. (a–c), shown are gene set enrichment analyses (a) and heat maps for the top 10 up‐regulated genes (b, c) for ATP BIOSYNTHETIC PROCESS and MICROTUBULE CYTOSKELETON ORGANIZATION in D + Q compared with vehicle group. (d, e), electron microscopy image of a vehicle and a D + Q treated VSMC, and summary of the results. Caveolae density, diameter of caveolae neck in VSMCs from vehicle‐ versus D + Q‐treated mice (four mice in each group). (f‐g), confocal immunofluorescence images and the line course of fluorescence changes. Immunofluorescently labeled with Ca V 3.2 (green) and Cav‐1 (red) in mesenteric arteries from vehicle‐ and D + Q‐treated mice. Bar, 20 μm. (h), Pearson's correlation coefficients for colocalization assays. The plot shows Pearson's correlation coefficients for the colocalization analysis ( n = 30–40 arteries). The Kruskal–Wallis H test was used for calculating statistical differences. Arteries were isolated from four mice in each group. (i), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle aged mouse and the time course of Ca 2+ fluorescence changes. (j), same as (h) but in the presence of Ni 2+ (50 μM). (k), same as (h) but in the cell from a D + Q treated aged mouse. (l), same as (j) but in the presence of Ni 2+ . (m–p), summary of the results. Ca 2+ spark frequency (m) and fraction of cells producing Ca 2+ sparks (n) in VSMCs from aged mice ( n = 124), in VSMCs from aged mice incubated with Ni 2+ ( n = 109), in VSMCs from D + Q‐treated aged mice ( n = 176), and in VSMCs from D + Q‐treated aged mice incubated with Ni 2+ ( n = 115). Ca 2+ spark frequency (o) and fraction of cells producing Ca 2+ sparks (p) in VSMCs from D + Q treated aged mice incubated with Ni 2+ ( n = 112), with methyl‐β‐cyclodextrin ( n = 108), as well as Ni 2+ +methyl‐β‐cyclodextrin ( n = 96). Cells were isolated from four mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05; n.s., not significant.
    Figure Legend Snippet: D + Q promote caveolae remodeling and Ca V 3.2‐Cav‐1 co‐localization to rescue caveolar Ca V 3.2‐RyR axis in aged VSMC. (a–c), shown are gene set enrichment analyses (a) and heat maps for the top 10 up‐regulated genes (b, c) for ATP BIOSYNTHETIC PROCESS and MICROTUBULE CYTOSKELETON ORGANIZATION in D + Q compared with vehicle group. (d, e), electron microscopy image of a vehicle and a D + Q treated VSMC, and summary of the results. Caveolae density, diameter of caveolae neck in VSMCs from vehicle‐ versus D + Q‐treated mice (four mice in each group). (f‐g), confocal immunofluorescence images and the line course of fluorescence changes. Immunofluorescently labeled with Ca V 3.2 (green) and Cav‐1 (red) in mesenteric arteries from vehicle‐ and D + Q‐treated mice. Bar, 20 μm. (h), Pearson's correlation coefficients for colocalization assays. The plot shows Pearson's correlation coefficients for the colocalization analysis ( n = 30–40 arteries). The Kruskal–Wallis H test was used for calculating statistical differences. Arteries were isolated from four mice in each group. (i), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle aged mouse and the time course of Ca 2+ fluorescence changes. (j), same as (h) but in the presence of Ni 2+ (50 μM). (k), same as (h) but in the cell from a D + Q treated aged mouse. (l), same as (j) but in the presence of Ni 2+ . (m–p), summary of the results. Ca 2+ spark frequency (m) and fraction of cells producing Ca 2+ sparks (n) in VSMCs from aged mice ( n = 124), in VSMCs from aged mice incubated with Ni 2+ ( n = 109), in VSMCs from D + Q‐treated aged mice ( n = 176), and in VSMCs from D + Q‐treated aged mice incubated with Ni 2+ ( n = 115). Ca 2+ spark frequency (o) and fraction of cells producing Ca 2+ sparks (p) in VSMCs from D + Q treated aged mice incubated with Ni 2+ ( n = 112), with methyl‐β‐cyclodextrin ( n = 108), as well as Ni 2+ +methyl‐β‐cyclodextrin ( n = 96). Cells were isolated from four mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05; n.s., not significant.

    Techniques Used: Electron Microscopy, Immunofluorescence, Fluorescence, Labeling, Isolation, Incubation

    cacna1c ca v 1 2  (Alomone Labs)


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    Alomone Labs cacna1c ca v 1 2
    Alteration of voltage-gated calcium channel gating properties in DM1 hiPSC-CMs. (A) Representative calcium currents. The dashed line represents zero current. The currents were obtained using 250-ms pulses from −40 mV to +60 mV in 5-mV increments. (B) Normalized intensity (I CaL )/voltage relationships (I/V) in CTRL ( N = 4, n = 16), DM1-1290 ( N = 5, n = 29), and DM1-1640 ( N = 4, n = 17) hiPSC-CMs. Calcium current densities were measured by normalizing current amplitudes to membrane capacitance. * p < 0.05 (CTRL vs. DM1-1290), and # p < 0.05, ##< 0.01 (CTRL vs. DM1-1640). The significance was determined using a two-way ANOVA with Šídák’s multiple comparisons test. (C) Steady state of activation and inactivation of I CaL currents. Inactivation currents were obtained using 30-ms test pulses to +10 mV after a 3,000-ms pre-pulse to potentials ranging from −40 mV to +20 mV. (D) Box and whiskers summarizing the half-activation and half-inactivation potentials. * p < 0.05, ** p < 0.01, and *** p < 0.001 as determined using a one-way ANOVA with Tukey’s multiple comparisons test. (E) qPCR analysis of <t>CACNA1C</t> and CACNA1D mRNA expression. CACNA1D is normalized to CACNA1C expression in CTRL ( N = 5, n = 7), DM1-1290 ( N = 6, n = 9), and DM1-1640 ( N = 5, n = 6) hiPSC-CMs. * p < 0.05, and ** p < 0.01 as determined using a non-parametric Kruskal-Wallis test with Dunn’s multiple comparisons test. (F) Western blot analysis of Ca V 1.2 channel expression. Box and whiskers showing the quantification of Ca V 1.2 channel expression normalized to the total protein for CTRL ( N = 4), DM1-1290 ( N = 5), and DM1-1640 hiPSC-CMs ( N = 5). * p < 0.05 as determined using a non-parametric Kruskal-Wallis test with Dunn’s multiple comparisons test. (G) Window current representing the overlap between the activation and inactivation curves. The replicate ( N ) corresponds to the number of differentiations while the replicate ( n ) corresponds to either the number of recorded cells or RNA or protein extract.
    Cacna1c Ca V 1 2, 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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    1) Product Images from "Cardiac involvement in patient-specific induced pluripotent stem cells of myotonic dystrophy type 1: unveiling the impact of voltage-gated sodium channels"

    Article Title: Cardiac involvement in patient-specific induced pluripotent stem cells of myotonic dystrophy type 1: unveiling the impact of voltage-gated sodium channels

    Journal: Frontiers in Physiology

    doi: 10.3389/fphys.2023.1258318

    Alteration of voltage-gated calcium channel gating properties in DM1 hiPSC-CMs. (A) Representative calcium currents. The dashed line represents zero current. The currents were obtained using 250-ms pulses from −40 mV to +60 mV in 5-mV increments. (B) Normalized intensity (I CaL )/voltage relationships (I/V) in CTRL ( N = 4, n = 16), DM1-1290 ( N = 5, n = 29), and DM1-1640 ( N = 4, n = 17) hiPSC-CMs. Calcium current densities were measured by normalizing current amplitudes to membrane capacitance. * p < 0.05 (CTRL vs. DM1-1290), and # p < 0.05, ##< 0.01 (CTRL vs. DM1-1640). The significance was determined using a two-way ANOVA with Šídák’s multiple comparisons test. (C) Steady state of activation and inactivation of I CaL currents. Inactivation currents were obtained using 30-ms test pulses to +10 mV after a 3,000-ms pre-pulse to potentials ranging from −40 mV to +20 mV. (D) Box and whiskers summarizing the half-activation and half-inactivation potentials. * p < 0.05, ** p < 0.01, and *** p < 0.001 as determined using a one-way ANOVA with Tukey’s multiple comparisons test. (E) qPCR analysis of CACNA1C and CACNA1D mRNA expression. CACNA1D is normalized to CACNA1C expression in CTRL ( N = 5, n = 7), DM1-1290 ( N = 6, n = 9), and DM1-1640 ( N = 5, n = 6) hiPSC-CMs. * p < 0.05, and ** p < 0.01 as determined using a non-parametric Kruskal-Wallis test with Dunn’s multiple comparisons test. (F) Western blot analysis of Ca V 1.2 channel expression. Box and whiskers showing the quantification of Ca V 1.2 channel expression normalized to the total protein for CTRL ( N = 4), DM1-1290 ( N = 5), and DM1-1640 hiPSC-CMs ( N = 5). * p < 0.05 as determined using a non-parametric Kruskal-Wallis test with Dunn’s multiple comparisons test. (G) Window current representing the overlap between the activation and inactivation curves. The replicate ( N ) corresponds to the number of differentiations while the replicate ( n ) corresponds to either the number of recorded cells or RNA or protein extract.
    Figure Legend Snippet: Alteration of voltage-gated calcium channel gating properties in DM1 hiPSC-CMs. (A) Representative calcium currents. The dashed line represents zero current. The currents were obtained using 250-ms pulses from −40 mV to +60 mV in 5-mV increments. (B) Normalized intensity (I CaL )/voltage relationships (I/V) in CTRL ( N = 4, n = 16), DM1-1290 ( N = 5, n = 29), and DM1-1640 ( N = 4, n = 17) hiPSC-CMs. Calcium current densities were measured by normalizing current amplitudes to membrane capacitance. * p < 0.05 (CTRL vs. DM1-1290), and # p < 0.05, ##< 0.01 (CTRL vs. DM1-1640). The significance was determined using a two-way ANOVA with Šídák’s multiple comparisons test. (C) Steady state of activation and inactivation of I CaL currents. Inactivation currents were obtained using 30-ms test pulses to +10 mV after a 3,000-ms pre-pulse to potentials ranging from −40 mV to +20 mV. (D) Box and whiskers summarizing the half-activation and half-inactivation potentials. * p < 0.05, ** p < 0.01, and *** p < 0.001 as determined using a one-way ANOVA with Tukey’s multiple comparisons test. (E) qPCR analysis of CACNA1C and CACNA1D mRNA expression. CACNA1D is normalized to CACNA1C expression in CTRL ( N = 5, n = 7), DM1-1290 ( N = 6, n = 9), and DM1-1640 ( N = 5, n = 6) hiPSC-CMs. * p < 0.05, and ** p < 0.01 as determined using a non-parametric Kruskal-Wallis test with Dunn’s multiple comparisons test. (F) Western blot analysis of Ca V 1.2 channel expression. Box and whiskers showing the quantification of Ca V 1.2 channel expression normalized to the total protein for CTRL ( N = 4), DM1-1290 ( N = 5), and DM1-1640 hiPSC-CMs ( N = 5). * p < 0.05 as determined using a non-parametric Kruskal-Wallis test with Dunn’s multiple comparisons test. (G) Window current representing the overlap between the activation and inactivation curves. The replicate ( N ) corresponds to the number of differentiations while the replicate ( n ) corresponds to either the number of recorded cells or RNA or protein extract.

    Techniques Used: Membrane, Activation Assay, Expressing, Western Blot

    rabbit anti ca v 1 2  (Alomone Labs)


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    Structured Review

    Alomone Labs rabbit anti ca v 1 2
    Different distribution of Ca V 1.2 channel and muscarinic M 4 receptor in the cell membrane in rat AMC cells. ( A ) Immunostaining of Ca V 1.2-YFP fusion protein expressed in HEK293T cells with a rabbit anti-Ca V 1.2 antibody (Ab). HEK293T transfected with a Ca V 1.2-YFP construct were labeled with the rabbit anti-Ca V 1.2 Ab. a and b represent confocal images of Ca V 1.2-like immunofluorescence and YFP fluorescence, respectively; c represents a differential interference contrast (DIC) image. The immunoreaction and YFP fluorescence were visualized with excitation at 514 nm and emission of 530–600 nm and with excitation at 633 and emission above 650 nm, respectively. ( B ) Fractionation analysis of rat adrenal medullae for integral membrane proteins. The cell membrane was divided into the raft and non-raft membrane domains by using discontinuous sucrose density gradient centrifugation (see the Materials and Methods). The same volume of each fraction with 5%–40% sucrose was immunoblotted for caveolin-1, transferrin receptor (R), muscarinic M 4 receptor, and TASK1 channel. Note that caveolin-1, a raft membrane marker, was enriched in the 20% fraction, whereas transferrin R, a non-raft membrane marker, was present in the 40% fraction. ( C ) Double staining for caveolin-1 and Ca V 1.2 and for M 4 receptor and Ca V 1.2 in rat AMC cells. The first column indicates confocal images of caveolin-1 and M 4 receptor-like immunofluorescence. The second column shows confocal images of Ca V 1.2-like immunofluorescence. The third column is a merge of immunofluorescence images. The fourth column shows DIC images. The calibration applies to all the images. Dissociated rat AMC cells were treated overnight with rabbit anti-Ca V 1.2 Ab (dilution, 1:50) and mouse anti-caveolin-1 Ab (1:20) or mouse anti-M 4 Ab (1:50). Ca V 1.2 and caveolin-1 or M 4 receptor-like immunoreactive material were visible as rhodamine and FITC-like fluorescence, respectively. ( D ) Summary of the coincidence rates of caveolin-1 (Cav1) and M 4 with Ca V 1.2. The data represent the mean ± SEM (Cav1/Ca V 1.2, n = 10; M 4 /Ca V 1.2, n = 5). Statistical significance was evaluated with an unpaired Student’s t test.
    Rabbit Anti Ca V 1 2, 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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    Images

    1) Product Images from "Muscarinic Receptor Stimulation Does Not Inhibit Voltage-dependent Ca 2+ Channels in Rat Adrenal Medullary Chromaffin Cells"

    Article Title: Muscarinic Receptor Stimulation Does Not Inhibit Voltage-dependent Ca 2+ Channels in Rat Adrenal Medullary Chromaffin Cells

    Journal: Acta Histochemica et Cytochemica

    doi: 10.1267/ahc.23-00042

    Different distribution of Ca V 1.2 channel and muscarinic M 4 receptor in the cell membrane in rat AMC cells. ( A ) Immunostaining of Ca V 1.2-YFP fusion protein expressed in HEK293T cells with a rabbit anti-Ca V 1.2 antibody (Ab). HEK293T transfected with a Ca V 1.2-YFP construct were labeled with the rabbit anti-Ca V 1.2 Ab. a and b represent confocal images of Ca V 1.2-like immunofluorescence and YFP fluorescence, respectively; c represents a differential interference contrast (DIC) image. The immunoreaction and YFP fluorescence were visualized with excitation at 514 nm and emission of 530–600 nm and with excitation at 633 and emission above 650 nm, respectively. ( B ) Fractionation analysis of rat adrenal medullae for integral membrane proteins. The cell membrane was divided into the raft and non-raft membrane domains by using discontinuous sucrose density gradient centrifugation (see the Materials and Methods). The same volume of each fraction with 5%–40% sucrose was immunoblotted for caveolin-1, transferrin receptor (R), muscarinic M 4 receptor, and TASK1 channel. Note that caveolin-1, a raft membrane marker, was enriched in the 20% fraction, whereas transferrin R, a non-raft membrane marker, was present in the 40% fraction. ( C ) Double staining for caveolin-1 and Ca V 1.2 and for M 4 receptor and Ca V 1.2 in rat AMC cells. The first column indicates confocal images of caveolin-1 and M 4 receptor-like immunofluorescence. The second column shows confocal images of Ca V 1.2-like immunofluorescence. The third column is a merge of immunofluorescence images. The fourth column shows DIC images. The calibration applies to all the images. Dissociated rat AMC cells were treated overnight with rabbit anti-Ca V 1.2 Ab (dilution, 1:50) and mouse anti-caveolin-1 Ab (1:20) or mouse anti-M 4 Ab (1:50). Ca V 1.2 and caveolin-1 or M 4 receptor-like immunoreactive material were visible as rhodamine and FITC-like fluorescence, respectively. ( D ) Summary of the coincidence rates of caveolin-1 (Cav1) and M 4 with Ca V 1.2. The data represent the mean ± SEM (Cav1/Ca V 1.2, n = 10; M 4 /Ca V 1.2, n = 5). Statistical significance was evaluated with an unpaired Student’s t test.
    Figure Legend Snippet: Different distribution of Ca V 1.2 channel and muscarinic M 4 receptor in the cell membrane in rat AMC cells. ( A ) Immunostaining of Ca V 1.2-YFP fusion protein expressed in HEK293T cells with a rabbit anti-Ca V 1.2 antibody (Ab). HEK293T transfected with a Ca V 1.2-YFP construct were labeled with the rabbit anti-Ca V 1.2 Ab. a and b represent confocal images of Ca V 1.2-like immunofluorescence and YFP fluorescence, respectively; c represents a differential interference contrast (DIC) image. The immunoreaction and YFP fluorescence were visualized with excitation at 514 nm and emission of 530–600 nm and with excitation at 633 and emission above 650 nm, respectively. ( B ) Fractionation analysis of rat adrenal medullae for integral membrane proteins. The cell membrane was divided into the raft and non-raft membrane domains by using discontinuous sucrose density gradient centrifugation (see the Materials and Methods). The same volume of each fraction with 5%–40% sucrose was immunoblotted for caveolin-1, transferrin receptor (R), muscarinic M 4 receptor, and TASK1 channel. Note that caveolin-1, a raft membrane marker, was enriched in the 20% fraction, whereas transferrin R, a non-raft membrane marker, was present in the 40% fraction. ( C ) Double staining for caveolin-1 and Ca V 1.2 and for M 4 receptor and Ca V 1.2 in rat AMC cells. The first column indicates confocal images of caveolin-1 and M 4 receptor-like immunofluorescence. The second column shows confocal images of Ca V 1.2-like immunofluorescence. The third column is a merge of immunofluorescence images. The fourth column shows DIC images. The calibration applies to all the images. Dissociated rat AMC cells were treated overnight with rabbit anti-Ca V 1.2 Ab (dilution, 1:50) and mouse anti-caveolin-1 Ab (1:20) or mouse anti-M 4 Ab (1:50). Ca V 1.2 and caveolin-1 or M 4 receptor-like immunoreactive material were visible as rhodamine and FITC-like fluorescence, respectively. ( D ) Summary of the coincidence rates of caveolin-1 (Cav1) and M 4 with Ca V 1.2. The data represent the mean ± SEM (Cav1/Ca V 1.2, n = 10; M 4 /Ca V 1.2, n = 5). Statistical significance was evaluated with an unpaired Student’s t test.

    Techniques Used: Membrane, Immunostaining, Transfection, Construct, Labeling, Immunofluorescence, Fluorescence, Fractionation, Gradient Centrifugation, Marker, Double Staining

    Diagram showing localization of caveolin-1, Ca V 1.2, muscarinic M 4 receptor subtype, and TASK1 in the raft and non-raft membrane domains.
    Figure Legend Snippet: Diagram showing localization of caveolin-1, Ca V 1.2, muscarinic M 4 receptor subtype, and TASK1 in the raft and non-raft membrane domains.

    Techniques Used: Membrane

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    Alomone Labs ca v 1 2
    Primary antibodies.
    Ca V 1 2, 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 rabbit polyclonal igg anti ca v 1 2
    a Single-molecule localization microscopy (SMLM) map showing Ca V 1.2 channel localization and distribution in the t-tubules of young and old ventricular myocytes with or without ISO stimulation. Yellow boxes indicate the location of the regions of interest magnified in the top right of each image. b Dot-plots showing mean Ca V 1.2 channel cluster areas in young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 4, n = 11; ISO: N = 4, n = 10) myocytes. c , d show the same for RyR2 in young (control: N = 3, n = 15; ISO: N = 3, n = 15) and old (control: N = 3, n = 12; ISO: N = 3, n = 11) myocytes. Statistical analyses on data summarized in ( b , d ) were performed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in b and d are from NIA young. Data are presented as mean ± SEM. Source data are provided in the Source Data file.
    Rabbit Polyclonal Igg Anti Ca V 1 2, 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 anti ca v 1 2 antibody
    The protection of Ahf-caltide on the Langerdoff-perfused rat heart subjected to H 2 O 2 treatment. ( A ) Schematic diagram of the experimental protocol. ( B , C ) Time course of changes in heart rate ( B ) and coronary flow ( C ) among groups. ( D ) LDH activity of coronary effluent in groups. ( E , F ) Representatives immunoblots ( E ) and quantitative analysis ( F ) of Ca V 1.2, Ca V β2, CAST and BNP levels in hearts of groups. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. control group; # p < 0.05 and ## p < 0.01 vs. H 2 O 2 group. NS, not significant. Results represent at least five independent experiments.
    Anti Ca V 1 2 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
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    86
    Alomone Labs anti ca v 3 2 rabbit
    D + Q turnover caveolar Ca 2+ sparks in aged mesenteric VSMCs. (a), experimental design of senolytics administration is shown. (D, dasatinib; Q, quercetin; PEG, polyethylene glycol). (b), heat map of RNA‐seq data of mesenteric artery from young, vehicle‐, and D + Q‐treatment mice ( n = 4 samples for each group). (c–f) Shown are gene set enrichment analyses (c, d) and heat maps for the top 20 up‐regulated genes for CELL CYCLE (e) and APOPTOSIS (f) in D + Q compared with vehicle group. FDR, false discovery rate. (g), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle‐aged mouse and the time course of Ca 2+ fluorescence changes. (h), same as (g) but in the presence of methyl‐β‐cyclodextrin (dextrin, 10 mM, 90 min at room temperature). (i), same as (g) but in the cell from a D + Q‐treated aged mouse. (j), same as (i) but in the presence of methyl‐β‐cyclodextrin. (k–l), summary of the results. Ca 2+ spark frequency (e) and fraction of cells producing Ca 2+ sparks (f) in VSMCs from aged mice ( n = 134), in VSMCs from aged mice incubated with methyl‐β‐cyclodextrin ( n = 95), in VSMCs from D + Q treated aged mice ( n = 175), and in VSMCs from D + Q treated aged mice incubated with methyl‐β‐cyclodextrin ( n = 175). Cells were isolated from 4 mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05 ; n.s., not significant. (m), western blot analysis of Ca V 3.2, Caveolin‐1 proteins in mesenteric arteries of aged versus D + Q mice. (n), quantification of western blot results. Western blot results were analyzed from 8 mice in each group. * p < 0.05 ; n.s., not significant; Cav‐1, Caveolin‐1.
    Anti Ca V 3 2 Rabbit, 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 cacna1c ca v 1 2
    Alteration of voltage-gated calcium channel gating properties in DM1 hiPSC-CMs. (A) Representative calcium currents. The dashed line represents zero current. The currents were obtained using 250-ms pulses from −40 mV to +60 mV in 5-mV increments. (B) Normalized intensity (I CaL )/voltage relationships (I/V) in CTRL ( N = 4, n = 16), DM1-1290 ( N = 5, n = 29), and DM1-1640 ( N = 4, n = 17) hiPSC-CMs. Calcium current densities were measured by normalizing current amplitudes to membrane capacitance. * p < 0.05 (CTRL vs. DM1-1290), and # p < 0.05, ##< 0.01 (CTRL vs. DM1-1640). The significance was determined using a two-way ANOVA with Šídák’s multiple comparisons test. (C) Steady state of activation and inactivation of I CaL currents. Inactivation currents were obtained using 30-ms test pulses to +10 mV after a 3,000-ms pre-pulse to potentials ranging from −40 mV to +20 mV. (D) Box and whiskers summarizing the half-activation and half-inactivation potentials. * p < 0.05, ** p < 0.01, and *** p < 0.001 as determined using a one-way ANOVA with Tukey’s multiple comparisons test. (E) qPCR analysis of <t>CACNA1C</t> and CACNA1D mRNA expression. CACNA1D is normalized to CACNA1C expression in CTRL ( N = 5, n = 7), DM1-1290 ( N = 6, n = 9), and DM1-1640 ( N = 5, n = 6) hiPSC-CMs. * p < 0.05, and ** p < 0.01 as determined using a non-parametric Kruskal-Wallis test with Dunn’s multiple comparisons test. (F) Western blot analysis of Ca V 1.2 channel expression. Box and whiskers showing the quantification of Ca V 1.2 channel expression normalized to the total protein for CTRL ( N = 4), DM1-1290 ( N = 5), and DM1-1640 hiPSC-CMs ( N = 5). * p < 0.05 as determined using a non-parametric Kruskal-Wallis test with Dunn’s multiple comparisons test. (G) Window current representing the overlap between the activation and inactivation curves. The replicate ( N ) corresponds to the number of differentiations while the replicate ( n ) corresponds to either the number of recorded cells or RNA or protein extract.
    Cacna1c Ca V 1 2, 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 rabbit anti ca v 1 2
    Different distribution of Ca V 1.2 channel and muscarinic M 4 receptor in the cell membrane in rat AMC cells. ( A ) Immunostaining of Ca V 1.2-YFP fusion protein expressed in HEK293T cells with a rabbit anti-Ca V 1.2 antibody (Ab). HEK293T transfected with a Ca V 1.2-YFP construct were labeled with the rabbit anti-Ca V 1.2 Ab. a and b represent confocal images of Ca V 1.2-like immunofluorescence and YFP fluorescence, respectively; c represents a differential interference contrast (DIC) image. The immunoreaction and YFP fluorescence were visualized with excitation at 514 nm and emission of 530–600 nm and with excitation at 633 and emission above 650 nm, respectively. ( B ) Fractionation analysis of rat adrenal medullae for integral membrane proteins. The cell membrane was divided into the raft and non-raft membrane domains by using discontinuous sucrose density gradient centrifugation (see the Materials and Methods). The same volume of each fraction with 5%–40% sucrose was immunoblotted for caveolin-1, transferrin receptor (R), muscarinic M 4 receptor, and TASK1 channel. Note that caveolin-1, a raft membrane marker, was enriched in the 20% fraction, whereas transferrin R, a non-raft membrane marker, was present in the 40% fraction. ( C ) Double staining for caveolin-1 and Ca V 1.2 and for M 4 receptor and Ca V 1.2 in rat AMC cells. The first column indicates confocal images of caveolin-1 and M 4 receptor-like immunofluorescence. The second column shows confocal images of Ca V 1.2-like immunofluorescence. The third column is a merge of immunofluorescence images. The fourth column shows DIC images. The calibration applies to all the images. Dissociated rat AMC cells were treated overnight with rabbit anti-Ca V 1.2 Ab (dilution, 1:50) and mouse anti-caveolin-1 Ab (1:20) or mouse anti-M 4 Ab (1:50). Ca V 1.2 and caveolin-1 or M 4 receptor-like immunoreactive material were visible as rhodamine and FITC-like fluorescence, respectively. ( D ) Summary of the coincidence rates of caveolin-1 (Cav1) and M 4 with Ca V 1.2. The data represent the mean ± SEM (Cav1/Ca V 1.2, n = 10; M 4 /Ca V 1.2, n = 5). Statistical significance was evaluated with an unpaired Student’s t test.
    Rabbit Anti Ca V 1 2, 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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    Image Search Results


    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: MS , Ca v 1.2 , Rabbit , polyclonal , Alomone , ACC-003 , AB_2039771 , ND.

    Techniques: Concentration Assay

    a Single-molecule localization microscopy (SMLM) map showing Ca V 1.2 channel localization and distribution in the t-tubules of young and old ventricular myocytes with or without ISO stimulation. Yellow boxes indicate the location of the regions of interest magnified in the top right of each image. b Dot-plots showing mean Ca V 1.2 channel cluster areas in young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 4, n = 11; ISO: N = 4, n = 10) myocytes. c , d show the same for RyR2 in young (control: N = 3, n = 15; ISO: N = 3, n = 15) and old (control: N = 3, n = 12; ISO: N = 3, n = 11) myocytes. Statistical analyses on data summarized in ( b , d ) were performed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in b and d are from NIA young. Data are presented as mean ± SEM. Source data are provided in the Source Data file.

    Journal: Nature Communications

    Article Title: BIN1 knockdown rescues systolic dysfunction in aging male mouse hearts

    doi: 10.1038/s41467-024-47847-8

    Figure Lengend Snippet: a Single-molecule localization microscopy (SMLM) map showing Ca V 1.2 channel localization and distribution in the t-tubules of young and old ventricular myocytes with or without ISO stimulation. Yellow boxes indicate the location of the regions of interest magnified in the top right of each image. b Dot-plots showing mean Ca V 1.2 channel cluster areas in young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 4, n = 11; ISO: N = 4, n = 10) myocytes. c , d show the same for RyR2 in young (control: N = 3, n = 15; ISO: N = 3, n = 15) and old (control: N = 3, n = 12; ISO: N = 3, n = 11) myocytes. Statistical analyses on data summarized in ( b , d ) were performed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in b and d are from NIA young. Data are presented as mean ± SEM. Source data are provided in the Source Data file.

    Article Snippet: Cells were incubated overnight at 4 °C in rabbit polyclonal IgG anti-Ca V 1.2 (CACNA1C, ACC-003, Alomone Labs, Jerusalem, Israel; 1:300 dilution) or mouse monoclonal IgG1 anti-RyR2 (C3-33, MA3-916, Invitrogen, Waltham, MA, USA; 1:50 dilution) in blocking buffer.

    Techniques: Microscopy, Comparison

    a Airyscan super-resolution images of Ca V 1.2 (green) and EEA1 (magenta) immunostained myocytes with and without ISO. Bottom: Binary colocalization maps show pixels in which Ca V 1.2 and EEA1 completely overlapped. b dot-plots summarizing % colocalization between EEA1 and Ca V 1.2 young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 3, n = 14; ISO: N = 3, n = 14) myocytes, and c EEA1-positive endosome areas in young (control: N = 3, n = 15; ISO: N = 3, n = 16) and old (control: N = 3, n = 14; ISO: N = 3, n = 13) myocytes. Data were analyzed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in ( b , c ) are from JAX young. Note no significant differences in EEA1/Ca V 1.2 colocalization, responsivity to ISO, or endosome size was detected when young JAX and young NIA myocytes were compared (Supplementary Fig.  ). Data are presented as mean ± SEM. Source data are provided in the Source Data file.

    Journal: Nature Communications

    Article Title: BIN1 knockdown rescues systolic dysfunction in aging male mouse hearts

    doi: 10.1038/s41467-024-47847-8

    Figure Lengend Snippet: a Airyscan super-resolution images of Ca V 1.2 (green) and EEA1 (magenta) immunostained myocytes with and without ISO. Bottom: Binary colocalization maps show pixels in which Ca V 1.2 and EEA1 completely overlapped. b dot-plots summarizing % colocalization between EEA1 and Ca V 1.2 young (control: N = 3, n = 16; ISO: N = 3, n = 16) and old (control: N = 3, n = 14; ISO: N = 3, n = 14) myocytes, and c EEA1-positive endosome areas in young (control: N = 3, n = 15; ISO: N = 3, n = 16) and old (control: N = 3, n = 14; ISO: N = 3, n = 13) myocytes. Data were analyzed using two-way ANOVAs with multiple comparison post-hoc tests. Young data in ( b , c ) are from JAX young. Note no significant differences in EEA1/Ca V 1.2 colocalization, responsivity to ISO, or endosome size was detected when young JAX and young NIA myocytes were compared (Supplementary Fig. ). Data are presented as mean ± SEM. Source data are provided in the Source Data file.

    Article Snippet: Cells were incubated overnight at 4 °C in rabbit polyclonal IgG anti-Ca V 1.2 (CACNA1C, ACC-003, Alomone Labs, Jerusalem, Israel; 1:300 dilution) or mouse monoclonal IgG1 anti-RyR2 (C3-33, MA3-916, Invitrogen, Waltham, MA, USA; 1:50 dilution) in blocking buffer.

    Techniques: Comparison

    a Representative whole-cell currents from shRNA-scrmb and shRNA-mBIN1 myocytes before and during ISO application. b fold change in peak I Ca with ISO in young, old, shRNA-scrmb ( N = 3, n = 7), and shRNA-mBIN1 ( N = 3, n = 9) myocytes. c Representative Ca 2+ transients recorded from old shRNA-scrmb and shRNA-mBIN1 myocytes before and after ISO.  Fold increase after ISO from young, old, shRNA-scrmb ( N = 3, n = 14), and shRNA-mBIN1 ( N = 5, n = 18) myocytes. e SMLM localization maps showing Ca V 1.2 channel localization on t-tubules of myocytes from old shRNA-scrmb and shRNA-mBIN1, with or without ISO stimulation. Regions of interest are highlighted by yellow boxes. f Fold change in mean Ca V 1.2 channel cluster area with ISO in the young, old, old shRNA-scrmb (control: N = 3, n = 14; ISO: N = 3, n = 13) and shRNA-mBIN1 myocytes (control: N = 3, n = 12; ISO: N = 3, n = 11). g , h show the same layout for RyR2 immunostained old shRNA-scrmb (control: N = 3, n = 12; ISO: N = 3, n = 13) and shRNA-mBIN1 myocytes (control: N = 3, n = 9; ISO: N = 3, n = 8). Old and young data points in ( b ,  , f , h ) are reproduced from data in Figs.  b, h, and  b,  respectively. Statistical analysis was performed on data in ( b ,  , f , h ) using one-way ANOVAs with multiple comparison post-hoc tests. Young data in ( b ) are pooled from NIA young and JAX young myocytes, data in (  , f , h ) are from NIA young myocytes. Note there was no significant difference in I Ca , Ca V 1.2, and RyR2 cluster areas when JAX and NIA young mice were compared (see Supplementary Fig.  ). Data are presented as mean ± SEM. Source data are provided in the Source Data file.

    Journal: Nature Communications

    Article Title: BIN1 knockdown rescues systolic dysfunction in aging male mouse hearts

    doi: 10.1038/s41467-024-47847-8

    Figure Lengend Snippet: a Representative whole-cell currents from shRNA-scrmb and shRNA-mBIN1 myocytes before and during ISO application. b fold change in peak I Ca with ISO in young, old, shRNA-scrmb ( N = 3, n = 7), and shRNA-mBIN1 ( N = 3, n = 9) myocytes. c Representative Ca 2+ transients recorded from old shRNA-scrmb and shRNA-mBIN1 myocytes before and after ISO. Fold increase after ISO from young, old, shRNA-scrmb ( N = 3, n = 14), and shRNA-mBIN1 ( N = 5, n = 18) myocytes. e SMLM localization maps showing Ca V 1.2 channel localization on t-tubules of myocytes from old shRNA-scrmb and shRNA-mBIN1, with or without ISO stimulation. Regions of interest are highlighted by yellow boxes. f Fold change in mean Ca V 1.2 channel cluster area with ISO in the young, old, old shRNA-scrmb (control: N = 3, n = 14; ISO: N = 3, n = 13) and shRNA-mBIN1 myocytes (control: N = 3, n = 12; ISO: N = 3, n = 11). g , h show the same layout for RyR2 immunostained old shRNA-scrmb (control: N = 3, n = 12; ISO: N = 3, n = 13) and shRNA-mBIN1 myocytes (control: N = 3, n = 9; ISO: N = 3, n = 8). Old and young data points in ( b , , f , h ) are reproduced from data in Figs. b, h, and b, respectively. Statistical analysis was performed on data in ( b , , f , h ) using one-way ANOVAs with multiple comparison post-hoc tests. Young data in ( b ) are pooled from NIA young and JAX young myocytes, data in ( , f , h ) are from NIA young myocytes. Note there was no significant difference in I Ca , Ca V 1.2, and RyR2 cluster areas when JAX and NIA young mice were compared (see Supplementary Fig. ). Data are presented as mean ± SEM. Source data are provided in the Source Data file.

    Article Snippet: Cells were incubated overnight at 4 °C in rabbit polyclonal IgG anti-Ca V 1.2 (CACNA1C, ACC-003, Alomone Labs, Jerusalem, Israel; 1:300 dilution) or mouse monoclonal IgG1 anti-RyR2 (C3-33, MA3-916, Invitrogen, Waltham, MA, USA; 1:50 dilution) in blocking buffer.

    Techniques: shRNA, Comparison

    The main findings of our study graphically illustrated and summarized. Top : In healthy young cells, Ca V 1.2 channels undergo endosomal recycling, where channels on endosomes are either marked for degradation through the late endosome pathway or are recycled to the sarcolemma through the fast and slow recycling pathways. Following β -adrenergic receptor ( β -AR) stimulation, a pool of channels localized to endosomes are mobilized to the membrane, resulting in larger Ca V 1.2 clusters along t-tubules. Across the dyad, RyR2 clusters on the sarcoplasmic reticulum also increase following βAR stimulation ensuring efficient Ca 2+ -induced Ca 2+ -release. This increase in cytosolic Ca 2+ , along with increased phosphorylation of cardiac Troponin I (cTnI) and cardiac myosin binding protein-C (cMyBP-C) within the sarcomere, allows for enhanced contractility under acute stress to cope with elevated hemodynamic and metabolic demands. Bottom : In aging, Bridging Integrator 1 (BIN1) protein levels are increased, accompanied by a swelling of endosomes and subsequent dysregulation of endosomal trafficking of Ca V 1.2. Ca V 1.2 and RyR2 channels are basally super-clustered at the dyads and lose β -AR responsivity. Reduced phosphorylation of cTnI and cMyBP-C result in systolic and diastolic dysfunction. BIN1 knockdown in aging recovers RyR2 clustering plasticity and Ca 2+ transient responsivity to β- AR stimulation. Phosphorylation of cMyBP-C is basally restored, and contractility is recovered to youthful levels. Thus, BIN1 knockdown rejuvenates the aging heart. Created with BioRender.com.

    Journal: Nature Communications

    Article Title: BIN1 knockdown rescues systolic dysfunction in aging male mouse hearts

    doi: 10.1038/s41467-024-47847-8

    Figure Lengend Snippet: The main findings of our study graphically illustrated and summarized. Top : In healthy young cells, Ca V 1.2 channels undergo endosomal recycling, where channels on endosomes are either marked for degradation through the late endosome pathway or are recycled to the sarcolemma through the fast and slow recycling pathways. Following β -adrenergic receptor ( β -AR) stimulation, a pool of channels localized to endosomes are mobilized to the membrane, resulting in larger Ca V 1.2 clusters along t-tubules. Across the dyad, RyR2 clusters on the sarcoplasmic reticulum also increase following βAR stimulation ensuring efficient Ca 2+ -induced Ca 2+ -release. This increase in cytosolic Ca 2+ , along with increased phosphorylation of cardiac Troponin I (cTnI) and cardiac myosin binding protein-C (cMyBP-C) within the sarcomere, allows for enhanced contractility under acute stress to cope with elevated hemodynamic and metabolic demands. Bottom : In aging, Bridging Integrator 1 (BIN1) protein levels are increased, accompanied by a swelling of endosomes and subsequent dysregulation of endosomal trafficking of Ca V 1.2. Ca V 1.2 and RyR2 channels are basally super-clustered at the dyads and lose β -AR responsivity. Reduced phosphorylation of cTnI and cMyBP-C result in systolic and diastolic dysfunction. BIN1 knockdown in aging recovers RyR2 clustering plasticity and Ca 2+ transient responsivity to β- AR stimulation. Phosphorylation of cMyBP-C is basally restored, and contractility is recovered to youthful levels. Thus, BIN1 knockdown rejuvenates the aging heart. Created with BioRender.com.

    Article Snippet: Cells were incubated overnight at 4 °C in rabbit polyclonal IgG anti-Ca V 1.2 (CACNA1C, ACC-003, Alomone Labs, Jerusalem, Israel; 1:300 dilution) or mouse monoclonal IgG1 anti-RyR2 (C3-33, MA3-916, Invitrogen, Waltham, MA, USA; 1:50 dilution) in blocking buffer.

    Techniques: Membrane, Binding Assay

    The protection of Ahf-caltide on the Langerdoff-perfused rat heart subjected to H 2 O 2 treatment. ( A ) Schematic diagram of the experimental protocol. ( B , C ) Time course of changes in heart rate ( B ) and coronary flow ( C ) among groups. ( D ) LDH activity of coronary effluent in groups. ( E , F ) Representatives immunoblots ( E ) and quantitative analysis ( F ) of Ca V 1.2, Ca V β2, CAST and BNP levels in hearts of groups. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. control group; # p < 0.05 and ## p < 0.01 vs. H 2 O 2 group. NS, not significant. Results represent at least five independent experiments.

    Journal: International Journal of Molecular Sciences

    Article Title: Ahf-Caltide, a Novel Polypeptide Derived from Calpastatin, Protects against Oxidative Stress Injury by Stabilizing the Expression of Ca V 1.2 Calcium Channel

    doi: 10.3390/ijms242115729

    Figure Lengend Snippet: The protection of Ahf-caltide on the Langerdoff-perfused rat heart subjected to H 2 O 2 treatment. ( A ) Schematic diagram of the experimental protocol. ( B , C ) Time course of changes in heart rate ( B ) and coronary flow ( C ) among groups. ( D ) LDH activity of coronary effluent in groups. ( E , F ) Representatives immunoblots ( E ) and quantitative analysis ( F ) of Ca V 1.2, Ca V β2, CAST and BNP levels in hearts of groups. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. control group; # p < 0.05 and ## p < 0.01 vs. H 2 O 2 group. NS, not significant. Results represent at least five independent experiments.

    Article Snippet: Anti-BNP antibody was purchased from Proteintech (Shanghai, China, Ag28102), anti-CAST antibody from Cell Signaling Technology (Danvers, MA, USA, #4146), anti-Ca V 1.2 antibody from Alomone labs (Jerusalem, Israel, ACC-003), anti-Ca V β2 antibody from Abcam (Cambridge, UK, ab 139528) and anti-GAPDH antibody was purchased from TransGen Biotech (Beijing, China, HC301).

    Techniques: Activity Assay, Western Blot

    The protection of Ahf-caltide on the Langerdoff-perfused rat hearts subjected to IR injury. ( A ) Schematic diagram of the experimental protocol. ( B , C ) Time course of changes in heart rate ( B ) and coronary flow ( C ) among groups. ( D ) LDH activity of coronary effluent in groups. ( E , F ) The SOD activity ( E ) and MDA content ( F ) in the heart tissues. ( G , H ) Representative images ( G ) and quantitative analysis ( H ) of Ca V 1.2, Ca V β2 and CAST in hearts among groups. * p < 0.05 and *** p < 0.001 vs. control group; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. IR group. NS, not significant. Results represent at least five independent experiments.

    Journal: International Journal of Molecular Sciences

    Article Title: Ahf-Caltide, a Novel Polypeptide Derived from Calpastatin, Protects against Oxidative Stress Injury by Stabilizing the Expression of Ca V 1.2 Calcium Channel

    doi: 10.3390/ijms242115729

    Figure Lengend Snippet: The protection of Ahf-caltide on the Langerdoff-perfused rat hearts subjected to IR injury. ( A ) Schematic diagram of the experimental protocol. ( B , C ) Time course of changes in heart rate ( B ) and coronary flow ( C ) among groups. ( D ) LDH activity of coronary effluent in groups. ( E , F ) The SOD activity ( E ) and MDA content ( F ) in the heart tissues. ( G , H ) Representative images ( G ) and quantitative analysis ( H ) of Ca V 1.2, Ca V β2 and CAST in hearts among groups. * p < 0.05 and *** p < 0.001 vs. control group; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. IR group. NS, not significant. Results represent at least five independent experiments.

    Article Snippet: Anti-BNP antibody was purchased from Proteintech (Shanghai, China, Ag28102), anti-CAST antibody from Cell Signaling Technology (Danvers, MA, USA, #4146), anti-Ca V 1.2 antibody from Alomone labs (Jerusalem, Israel, ACC-003), anti-Ca V β2 antibody from Abcam (Cambridge, UK, ab 139528) and anti-GAPDH antibody was purchased from TransGen Biotech (Beijing, China, HC301).

    Techniques: Activity Assay

    The protection by Ahf-caltide on H9c2 cell injury induced by CoCl 2 treatment. ( A ) The localization of Ahf-caltide in H9c2 cells examined by fluorescence microscopy (scale bars, 100 μm). ( B ) The cell viability was detected by CCK-8. ( C ) LDH activity of supernatant in groups. ( D , E ) ROS level in groups (scale bars, 100 μm). ( F , G ) Representative immunoblots ( F ) and quantitative analysis ( G ) of expression levels of Ca V 1.2, Ca V β2, CAST in groups. ( H , I ) Representative immunoblots ( H ) and quantitative analysis ( I ) of the effects of cycloheximide (CHX) on the expression of Ca V 1.2, Ca V β2, CAST in groups. ( J , K ) Representative immunoblots ( J ) and quantitative analysis ( K ) of the effects of MG-132/Leupeptin on the levels of Ca V 1.2, Ca V β2, CAST of groups. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. control group or as indicated on the figure; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. CoCl 2 group. NS, no significance. Results represent at least three independent experiments.

    Journal: International Journal of Molecular Sciences

    Article Title: Ahf-Caltide, a Novel Polypeptide Derived from Calpastatin, Protects against Oxidative Stress Injury by Stabilizing the Expression of Ca V 1.2 Calcium Channel

    doi: 10.3390/ijms242115729

    Figure Lengend Snippet: The protection by Ahf-caltide on H9c2 cell injury induced by CoCl 2 treatment. ( A ) The localization of Ahf-caltide in H9c2 cells examined by fluorescence microscopy (scale bars, 100 μm). ( B ) The cell viability was detected by CCK-8. ( C ) LDH activity of supernatant in groups. ( D , E ) ROS level in groups (scale bars, 100 μm). ( F , G ) Representative immunoblots ( F ) and quantitative analysis ( G ) of expression levels of Ca V 1.2, Ca V β2, CAST in groups. ( H , I ) Representative immunoblots ( H ) and quantitative analysis ( I ) of the effects of cycloheximide (CHX) on the expression of Ca V 1.2, Ca V β2, CAST in groups. ( J , K ) Representative immunoblots ( J ) and quantitative analysis ( K ) of the effects of MG-132/Leupeptin on the levels of Ca V 1.2, Ca V β2, CAST of groups. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. control group or as indicated on the figure; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. CoCl 2 group. NS, no significance. Results represent at least three independent experiments.

    Article Snippet: Anti-BNP antibody was purchased from Proteintech (Shanghai, China, Ag28102), anti-CAST antibody from Cell Signaling Technology (Danvers, MA, USA, #4146), anti-Ca V 1.2 antibody from Alomone labs (Jerusalem, Israel, ACC-003), anti-Ca V β2 antibody from Abcam (Cambridge, UK, ab 139528) and anti-GAPDH antibody was purchased from TransGen Biotech (Beijing, China, HC301).

    Techniques: Fluorescence, Microscopy, CCK-8 Assay, Activity Assay, Western Blot, Expressing

    Concentration-dependent effect of H 2 O 2 on CS L binding to N- and C-terminus of Ca V 1.2 channel α 1 subunit. GST pull-down assay for the effect of H 2 O 2 (10 −1 –10 −6 M) on the binding of CS L to CT1 ( A ) or PreIQ ( C ) or IQ ( E ) or NT ( K ). Plots of the effect of H 2 O 2 with fitted curves on the CS L binding to CT1 ( B ) or PreIQ ( D ) or IQ ( F ) or NT ( L ). Pull-down assay ( G ) and binding curves ( H ) for CS L binding to NT. Pull-down assay for the effect of [Ca 2+ ] ( I ) and quantitative analysis ( J ) on the binding of CS L to NT. Symbols are as indicated in each graph. NS, no significance. Protein bands were visualized using Coomassie brilliant blue staining and indicated by arrows. Results represent at least four independent experiments.

    Journal: International Journal of Molecular Sciences

    Article Title: Ahf-Caltide, a Novel Polypeptide Derived from Calpastatin, Protects against Oxidative Stress Injury by Stabilizing the Expression of Ca V 1.2 Calcium Channel

    doi: 10.3390/ijms242115729

    Figure Lengend Snippet: Concentration-dependent effect of H 2 O 2 on CS L binding to N- and C-terminus of Ca V 1.2 channel α 1 subunit. GST pull-down assay for the effect of H 2 O 2 (10 −1 –10 −6 M) on the binding of CS L to CT1 ( A ) or PreIQ ( C ) or IQ ( E ) or NT ( K ). Plots of the effect of H 2 O 2 with fitted curves on the CS L binding to CT1 ( B ) or PreIQ ( D ) or IQ ( F ) or NT ( L ). Pull-down assay ( G ) and binding curves ( H ) for CS L binding to NT. Pull-down assay for the effect of [Ca 2+ ] ( I ) and quantitative analysis ( J ) on the binding of CS L to NT. Symbols are as indicated in each graph. NS, no significance. Protein bands were visualized using Coomassie brilliant blue staining and indicated by arrows. Results represent at least four independent experiments.

    Article Snippet: Anti-BNP antibody was purchased from Proteintech (Shanghai, China, Ag28102), anti-CAST antibody from Cell Signaling Technology (Danvers, MA, USA, #4146), anti-Ca V 1.2 antibody from Alomone labs (Jerusalem, Israel, ACC-003), anti-Ca V β2 antibody from Abcam (Cambridge, UK, ab 139528) and anti-GAPDH antibody was purchased from TransGen Biotech (Beijing, China, HC301).

    Techniques: Concentration Assay, Binding Assay, Pull Down Assay, Staining

    Molecular docking analysis of CS L or CS L 54-64 binding to N- and C- terminus of Ca V 1.2 channel α 1 subunit. ( A – C ) Improved five groups of epitope clusters in  corresponding to the totally docked poses of ( A1 ) CS L /PreIQ, ( B1 ) CS L /IQ, ( C1 ) CS L /NT; the fingerprints for the interaction obtained from the five groups’ docked poses between CS L and ( A2 ) PreIQ, ( B2 ) IQ, ( C2 ) NT; the fingerprints for the interaction from the first group pose between CS L and ( A3 ) PreIQ, ( B3 ) IQ, ( C3 ) NT; the docked poses of CS L 54-64 binding to ( A4 ) PreIQ, ( B4 ) IQ, ( C4 ) NT; the interaction diagram of Lys 64, Lys 56 and Glu 54 on CS L 54-64 with PreIQ ( A5 ), IQ ( B5 ) and NT ( C5 ), respectively. The diagrams in the first group with the highest score are represented by blue “spaghetti” and the others with a translucent “spaghetti”. Blue structures represent PreIQ/IQ/NT, red structures represent CS L or CS L 54-64.

    Journal: International Journal of Molecular Sciences

    Article Title: Ahf-Caltide, a Novel Polypeptide Derived from Calpastatin, Protects against Oxidative Stress Injury by Stabilizing the Expression of Ca V 1.2 Calcium Channel

    doi: 10.3390/ijms242115729

    Figure Lengend Snippet: Molecular docking analysis of CS L or CS L 54-64 binding to N- and C- terminus of Ca V 1.2 channel α 1 subunit. ( A – C ) Improved five groups of epitope clusters in corresponding to the totally docked poses of ( A1 ) CS L /PreIQ, ( B1 ) CS L /IQ, ( C1 ) CS L /NT; the fingerprints for the interaction obtained from the five groups’ docked poses between CS L and ( A2 ) PreIQ, ( B2 ) IQ, ( C2 ) NT; the fingerprints for the interaction from the first group pose between CS L and ( A3 ) PreIQ, ( B3 ) IQ, ( C3 ) NT; the docked poses of CS L 54-64 binding to ( A4 ) PreIQ, ( B4 ) IQ, ( C4 ) NT; the interaction diagram of Lys 64, Lys 56 and Glu 54 on CS L 54-64 with PreIQ ( A5 ), IQ ( B5 ) and NT ( C5 ), respectively. The diagrams in the first group with the highest score are represented by blue “spaghetti” and the others with a translucent “spaghetti”. Blue structures represent PreIQ/IQ/NT, red structures represent CS L or CS L 54-64.

    Article Snippet: Anti-BNP antibody was purchased from Proteintech (Shanghai, China, Ag28102), anti-CAST antibody from Cell Signaling Technology (Danvers, MA, USA, #4146), anti-Ca V 1.2 antibody from Alomone labs (Jerusalem, Israel, ACC-003), anti-Ca V β2 antibody from Abcam (Cambridge, UK, ab 139528) and anti-GAPDH antibody was purchased from TransGen Biotech (Beijing, China, HC301).

    Techniques: Binding Assay

    Journal: International Journal of Molecular Sciences

    Article Title: Ahf-Caltide, a Novel Polypeptide Derived from Calpastatin, Protects against Oxidative Stress Injury by Stabilizing the Expression of Ca V 1.2 Calcium Channel

    doi: 10.3390/ijms242115729

    Figure Lengend Snippet: Predicted binding sites of CS L 54-65 to Ca V 1.2 channels.

    Article Snippet: Anti-BNP antibody was purchased from Proteintech (Shanghai, China, Ag28102), anti-CAST antibody from Cell Signaling Technology (Danvers, MA, USA, #4146), anti-Ca V 1.2 antibody from Alomone labs (Jerusalem, Israel, ACC-003), anti-Ca V β2 antibody from Abcam (Cambridge, UK, ab 139528) and anti-GAPDH antibody was purchased from TransGen Biotech (Beijing, China, HC301).

    Techniques: Binding Assay

    D + Q turnover caveolar Ca 2+ sparks in aged mesenteric VSMCs. (a), experimental design of senolytics administration is shown. (D, dasatinib; Q, quercetin; PEG, polyethylene glycol). (b), heat map of RNA‐seq data of mesenteric artery from young, vehicle‐, and D + Q‐treatment mice ( n = 4 samples for each group). (c–f) Shown are gene set enrichment analyses (c, d) and heat maps for the top 20 up‐regulated genes for CELL CYCLE (e) and APOPTOSIS (f) in D + Q compared with vehicle group. FDR, false discovery rate. (g), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle‐aged mouse and the time course of Ca 2+ fluorescence changes. (h), same as (g) but in the presence of methyl‐β‐cyclodextrin (dextrin, 10 mM, 90 min at room temperature). (i), same as (g) but in the cell from a D + Q‐treated aged mouse. (j), same as (i) but in the presence of methyl‐β‐cyclodextrin. (k–l), summary of the results. Ca 2+ spark frequency (e) and fraction of cells producing Ca 2+ sparks (f) in VSMCs from aged mice ( n = 134), in VSMCs from aged mice incubated with methyl‐β‐cyclodextrin ( n = 95), in VSMCs from D + Q treated aged mice ( n = 175), and in VSMCs from D + Q treated aged mice incubated with methyl‐β‐cyclodextrin ( n = 175). Cells were isolated from 4 mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05 ; n.s., not significant. (m), western blot analysis of Ca V 3.2, Caveolin‐1 proteins in mesenteric arteries of aged versus D + Q mice. (n), quantification of western blot results. Western blot results were analyzed from 8 mice in each group. * p < 0.05 ; n.s., not significant; Cav‐1, Caveolin‐1.

    Journal: Aging Cell

    Article Title: Senolytics prevent caveolar Ca V 3 . 2‐RyR axis malfunction in old vascular smooth muscle

    doi: 10.1111/acel.14002

    Figure Lengend Snippet: D + Q turnover caveolar Ca 2+ sparks in aged mesenteric VSMCs. (a), experimental design of senolytics administration is shown. (D, dasatinib; Q, quercetin; PEG, polyethylene glycol). (b), heat map of RNA‐seq data of mesenteric artery from young, vehicle‐, and D + Q‐treatment mice ( n = 4 samples for each group). (c–f) Shown are gene set enrichment analyses (c, d) and heat maps for the top 20 up‐regulated genes for CELL CYCLE (e) and APOPTOSIS (f) in D + Q compared with vehicle group. FDR, false discovery rate. (g), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle‐aged mouse and the time course of Ca 2+ fluorescence changes. (h), same as (g) but in the presence of methyl‐β‐cyclodextrin (dextrin, 10 mM, 90 min at room temperature). (i), same as (g) but in the cell from a D + Q‐treated aged mouse. (j), same as (i) but in the presence of methyl‐β‐cyclodextrin. (k–l), summary of the results. Ca 2+ spark frequency (e) and fraction of cells producing Ca 2+ sparks (f) in VSMCs from aged mice ( n = 134), in VSMCs from aged mice incubated with methyl‐β‐cyclodextrin ( n = 95), in VSMCs from D + Q treated aged mice ( n = 175), and in VSMCs from D + Q treated aged mice incubated with methyl‐β‐cyclodextrin ( n = 175). Cells were isolated from 4 mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05 ; n.s., not significant. (m), western blot analysis of Ca V 3.2, Caveolin‐1 proteins in mesenteric arteries of aged versus D + Q mice. (n), quantification of western blot results. Western blot results were analyzed from 8 mice in each group. * p < 0.05 ; n.s., not significant; Cav‐1, Caveolin‐1.

    Article Snippet: Antibodies: anti‐Ca V 3.2‐rabbit (Alomone Labs, #ACC‐025), anti‐caveolin‐1‐mouse (Beyotime, #AF0087), DAPI (Sigma, #D9542).

    Techniques: RNA Sequencing Assay, Fluorescence, Incubation, Isolation, Western Blot

    D + Q promote caveolae remodeling and Ca V 3.2‐Cav‐1 co‐localization to rescue caveolar Ca V 3.2‐RyR axis in aged VSMC. (a–c), shown are gene set enrichment analyses (a) and heat maps for the top 10 up‐regulated genes (b, c) for ATP BIOSYNTHETIC PROCESS and MICROTUBULE CYTOSKELETON ORGANIZATION in D + Q compared with vehicle group. (d, e), electron microscopy image of a vehicle and a D + Q treated VSMC, and summary of the results. Caveolae density, diameter of caveolae neck in VSMCs from vehicle‐ versus D + Q‐treated mice (four mice in each group). (f‐g), confocal immunofluorescence images and the line course of fluorescence changes. Immunofluorescently labeled with Ca V 3.2 (green) and Cav‐1 (red) in mesenteric arteries from vehicle‐ and D + Q‐treated mice. Bar, 20 μm. (h), Pearson's correlation coefficients for colocalization assays. The plot shows Pearson's correlation coefficients for the colocalization analysis ( n = 30–40 arteries). The Kruskal–Wallis H test was used for calculating statistical differences. Arteries were isolated from four mice in each group. (i), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle aged mouse and the time course of Ca 2+ fluorescence changes. (j), same as (h) but in the presence of Ni 2+ (50 μM). (k), same as (h) but in the cell from a D + Q treated aged mouse. (l), same as (j) but in the presence of Ni 2+ . (m–p), summary of the results. Ca 2+ spark frequency (m) and fraction of cells producing Ca 2+ sparks (n) in VSMCs from aged mice ( n = 124), in VSMCs from aged mice incubated with Ni 2+ ( n = 109), in VSMCs from D + Q‐treated aged mice ( n = 176), and in VSMCs from D + Q‐treated aged mice incubated with Ni 2+ ( n = 115). Ca 2+ spark frequency (o) and fraction of cells producing Ca 2+ sparks (p) in VSMCs from D + Q treated aged mice incubated with Ni 2+ ( n = 112), with methyl‐β‐cyclodextrin ( n = 108), as well as Ni 2+ +methyl‐β‐cyclodextrin ( n = 96). Cells were isolated from four mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05; n.s., not significant.

    Journal: Aging Cell

    Article Title: Senolytics prevent caveolar Ca V 3 . 2‐RyR axis malfunction in old vascular smooth muscle

    doi: 10.1111/acel.14002

    Figure Lengend Snippet: D + Q promote caveolae remodeling and Ca V 3.2‐Cav‐1 co‐localization to rescue caveolar Ca V 3.2‐RyR axis in aged VSMC. (a–c), shown are gene set enrichment analyses (a) and heat maps for the top 10 up‐regulated genes (b, c) for ATP BIOSYNTHETIC PROCESS and MICROTUBULE CYTOSKELETON ORGANIZATION in D + Q compared with vehicle group. (d, e), electron microscopy image of a vehicle and a D + Q treated VSMC, and summary of the results. Caveolae density, diameter of caveolae neck in VSMCs from vehicle‐ versus D + Q‐treated mice (four mice in each group). (f‐g), confocal immunofluorescence images and the line course of fluorescence changes. Immunofluorescently labeled with Ca V 3.2 (green) and Cav‐1 (red) in mesenteric arteries from vehicle‐ and D + Q‐treated mice. Bar, 20 μm. (h), Pearson's correlation coefficients for colocalization assays. The plot shows Pearson's correlation coefficients for the colocalization analysis ( n = 30–40 arteries). The Kruskal–Wallis H test was used for calculating statistical differences. Arteries were isolated from four mice in each group. (i), Ca 2+ fluorescence line scan images of a Fluo‐4‐AM–loaded VSMC from a middle aged mouse and the time course of Ca 2+ fluorescence changes. (j), same as (h) but in the presence of Ni 2+ (50 μM). (k), same as (h) but in the cell from a D + Q treated aged mouse. (l), same as (j) but in the presence of Ni 2+ . (m–p), summary of the results. Ca 2+ spark frequency (m) and fraction of cells producing Ca 2+ sparks (n) in VSMCs from aged mice ( n = 124), in VSMCs from aged mice incubated with Ni 2+ ( n = 109), in VSMCs from D + Q‐treated aged mice ( n = 176), and in VSMCs from D + Q‐treated aged mice incubated with Ni 2+ ( n = 115). Ca 2+ spark frequency (o) and fraction of cells producing Ca 2+ sparks (p) in VSMCs from D + Q treated aged mice incubated with Ni 2+ ( n = 112), with methyl‐β‐cyclodextrin ( n = 108), as well as Ni 2+ +methyl‐β‐cyclodextrin ( n = 96). Cells were isolated from four mice in each group. VSMC, vascular smooth muscle cell. * p < 0.05; n.s., not significant.

    Article Snippet: Antibodies: anti‐Ca V 3.2‐rabbit (Alomone Labs, #ACC‐025), anti‐caveolin‐1‐mouse (Beyotime, #AF0087), DAPI (Sigma, #D9542).

    Techniques: Electron Microscopy, Immunofluorescence, Fluorescence, Labeling, Isolation, Incubation

    Alteration of voltage-gated calcium channel gating properties in DM1 hiPSC-CMs. (A) Representative calcium currents. The dashed line represents zero current. The currents were obtained using 250-ms pulses from −40 mV to +60 mV in 5-mV increments. (B) Normalized intensity (I CaL )/voltage relationships (I/V) in CTRL ( N = 4, n = 16), DM1-1290 ( N = 5, n = 29), and DM1-1640 ( N = 4, n = 17) hiPSC-CMs. Calcium current densities were measured by normalizing current amplitudes to membrane capacitance. * p < 0.05 (CTRL vs. DM1-1290), and # p < 0.05, ##< 0.01 (CTRL vs. DM1-1640). The significance was determined using a two-way ANOVA with Šídák’s multiple comparisons test. (C) Steady state of activation and inactivation of I CaL currents. Inactivation currents were obtained using 30-ms test pulses to +10 mV after a 3,000-ms pre-pulse to potentials ranging from −40 mV to +20 mV. (D) Box and whiskers summarizing the half-activation and half-inactivation potentials. * p < 0.05, ** p < 0.01, and *** p < 0.001 as determined using a one-way ANOVA with Tukey’s multiple comparisons test. (E) qPCR analysis of CACNA1C and CACNA1D mRNA expression. CACNA1D is normalized to CACNA1C expression in CTRL ( N = 5, n = 7), DM1-1290 ( N = 6, n = 9), and DM1-1640 ( N = 5, n = 6) hiPSC-CMs. * p < 0.05, and ** p < 0.01 as determined using a non-parametric Kruskal-Wallis test with Dunn’s multiple comparisons test. (F) Western blot analysis of Ca V 1.2 channel expression. Box and whiskers showing the quantification of Ca V 1.2 channel expression normalized to the total protein for CTRL ( N = 4), DM1-1290 ( N = 5), and DM1-1640 hiPSC-CMs ( N = 5). * p < 0.05 as determined using a non-parametric Kruskal-Wallis test with Dunn’s multiple comparisons test. (G) Window current representing the overlap between the activation and inactivation curves. The replicate ( N ) corresponds to the number of differentiations while the replicate ( n ) corresponds to either the number of recorded cells or RNA or protein extract.

    Journal: Frontiers in Physiology

    Article Title: Cardiac involvement in patient-specific induced pluripotent stem cells of myotonic dystrophy type 1: unveiling the impact of voltage-gated sodium channels

    doi: 10.3389/fphys.2023.1258318

    Figure Lengend Snippet: Alteration of voltage-gated calcium channel gating properties in DM1 hiPSC-CMs. (A) Representative calcium currents. The dashed line represents zero current. The currents were obtained using 250-ms pulses from −40 mV to +60 mV in 5-mV increments. (B) Normalized intensity (I CaL )/voltage relationships (I/V) in CTRL ( N = 4, n = 16), DM1-1290 ( N = 5, n = 29), and DM1-1640 ( N = 4, n = 17) hiPSC-CMs. Calcium current densities were measured by normalizing current amplitudes to membrane capacitance. * p < 0.05 (CTRL vs. DM1-1290), and # p < 0.05, ##< 0.01 (CTRL vs. DM1-1640). The significance was determined using a two-way ANOVA with Šídák’s multiple comparisons test. (C) Steady state of activation and inactivation of I CaL currents. Inactivation currents were obtained using 30-ms test pulses to +10 mV after a 3,000-ms pre-pulse to potentials ranging from −40 mV to +20 mV. (D) Box and whiskers summarizing the half-activation and half-inactivation potentials. * p < 0.05, ** p < 0.01, and *** p < 0.001 as determined using a one-way ANOVA with Tukey’s multiple comparisons test. (E) qPCR analysis of CACNA1C and CACNA1D mRNA expression. CACNA1D is normalized to CACNA1C expression in CTRL ( N = 5, n = 7), DM1-1290 ( N = 6, n = 9), and DM1-1640 ( N = 5, n = 6) hiPSC-CMs. * p < 0.05, and ** p < 0.01 as determined using a non-parametric Kruskal-Wallis test with Dunn’s multiple comparisons test. (F) Western blot analysis of Ca V 1.2 channel expression. Box and whiskers showing the quantification of Ca V 1.2 channel expression normalized to the total protein for CTRL ( N = 4), DM1-1290 ( N = 5), and DM1-1640 hiPSC-CMs ( N = 5). * p < 0.05 as determined using a non-parametric Kruskal-Wallis test with Dunn’s multiple comparisons test. (G) Window current representing the overlap between the activation and inactivation curves. The replicate ( N ) corresponds to the number of differentiations while the replicate ( n ) corresponds to either the number of recorded cells or RNA or protein extract.

    Article Snippet: The PVDF membranes were blocked and were incubated with the following primary antibodies: rabbit anti-SCN5A (Na V 1.5, 1:200, Cat# ASC-005, Alomone Labs, RRID: AB_2040001), rabbit anti-calcium voltage-gated channel subunit alpha 1 C (CACNA1C/Ca V 1.2, 1:200, Cat# ACC-003, Alomone Labs, RRID:AB_2039771), rabbit anti-MBNL1 (1/500, Cat# NBP2-55165, Novus Biologicals), rabbit anti-GJA1 (Cx43, 1:5000, Cat# ab11370, Abcam, RRID: AB_297976), mouse anti-CELF1 (1:1000, Cat# SC-20003, Santa Cruz Biotechnology, RRID: AB_627319), and mouse anti-DMPK (1:750, cat# SC-134319, Santa Cruz Biotechnology, RRID: AB_2091375).

    Techniques: Membrane, Activation Assay, Expressing, Western Blot

    Different distribution of Ca V 1.2 channel and muscarinic M 4 receptor in the cell membrane in rat AMC cells. ( A ) Immunostaining of Ca V 1.2-YFP fusion protein expressed in HEK293T cells with a rabbit anti-Ca V 1.2 antibody (Ab). HEK293T transfected with a Ca V 1.2-YFP construct were labeled with the rabbit anti-Ca V 1.2 Ab. a and b represent confocal images of Ca V 1.2-like immunofluorescence and YFP fluorescence, respectively; c represents a differential interference contrast (DIC) image. The immunoreaction and YFP fluorescence were visualized with excitation at 514 nm and emission of 530–600 nm and with excitation at 633 and emission above 650 nm, respectively. ( B ) Fractionation analysis of rat adrenal medullae for integral membrane proteins. The cell membrane was divided into the raft and non-raft membrane domains by using discontinuous sucrose density gradient centrifugation (see the Materials and Methods). The same volume of each fraction with 5%–40% sucrose was immunoblotted for caveolin-1, transferrin receptor (R), muscarinic M 4 receptor, and TASK1 channel. Note that caveolin-1, a raft membrane marker, was enriched in the 20% fraction, whereas transferrin R, a non-raft membrane marker, was present in the 40% fraction. ( C ) Double staining for caveolin-1 and Ca V 1.2 and for M 4 receptor and Ca V 1.2 in rat AMC cells. The first column indicates confocal images of caveolin-1 and M 4 receptor-like immunofluorescence. The second column shows confocal images of Ca V 1.2-like immunofluorescence. The third column is a merge of immunofluorescence images. The fourth column shows DIC images. The calibration applies to all the images. Dissociated rat AMC cells were treated overnight with rabbit anti-Ca V 1.2 Ab (dilution, 1:50) and mouse anti-caveolin-1 Ab (1:20) or mouse anti-M 4 Ab (1:50). Ca V 1.2 and caveolin-1 or M 4 receptor-like immunoreactive material were visible as rhodamine and FITC-like fluorescence, respectively. ( D ) Summary of the coincidence rates of caveolin-1 (Cav1) and M 4 with Ca V 1.2. The data represent the mean ± SEM (Cav1/Ca V 1.2, n = 10; M 4 /Ca V 1.2, n = 5). Statistical significance was evaluated with an unpaired Student’s t test.

    Journal: Acta Histochemica et Cytochemica

    Article Title: Muscarinic Receptor Stimulation Does Not Inhibit Voltage-dependent Ca 2+ Channels in Rat Adrenal Medullary Chromaffin Cells

    doi: 10.1267/ahc.23-00042

    Figure Lengend Snippet: Different distribution of Ca V 1.2 channel and muscarinic M 4 receptor in the cell membrane in rat AMC cells. ( A ) Immunostaining of Ca V 1.2-YFP fusion protein expressed in HEK293T cells with a rabbit anti-Ca V 1.2 antibody (Ab). HEK293T transfected with a Ca V 1.2-YFP construct were labeled with the rabbit anti-Ca V 1.2 Ab. a and b represent confocal images of Ca V 1.2-like immunofluorescence and YFP fluorescence, respectively; c represents a differential interference contrast (DIC) image. The immunoreaction and YFP fluorescence were visualized with excitation at 514 nm and emission of 530–600 nm and with excitation at 633 and emission above 650 nm, respectively. ( B ) Fractionation analysis of rat adrenal medullae for integral membrane proteins. The cell membrane was divided into the raft and non-raft membrane domains by using discontinuous sucrose density gradient centrifugation (see the Materials and Methods). The same volume of each fraction with 5%–40% sucrose was immunoblotted for caveolin-1, transferrin receptor (R), muscarinic M 4 receptor, and TASK1 channel. Note that caveolin-1, a raft membrane marker, was enriched in the 20% fraction, whereas transferrin R, a non-raft membrane marker, was present in the 40% fraction. ( C ) Double staining for caveolin-1 and Ca V 1.2 and for M 4 receptor and Ca V 1.2 in rat AMC cells. The first column indicates confocal images of caveolin-1 and M 4 receptor-like immunofluorescence. The second column shows confocal images of Ca V 1.2-like immunofluorescence. The third column is a merge of immunofluorescence images. The fourth column shows DIC images. The calibration applies to all the images. Dissociated rat AMC cells were treated overnight with rabbit anti-Ca V 1.2 Ab (dilution, 1:50) and mouse anti-caveolin-1 Ab (1:20) or mouse anti-M 4 Ab (1:50). Ca V 1.2 and caveolin-1 or M 4 receptor-like immunoreactive material were visible as rhodamine and FITC-like fluorescence, respectively. ( D ) Summary of the coincidence rates of caveolin-1 (Cav1) and M 4 with Ca V 1.2. The data represent the mean ± SEM (Cav1/Ca V 1.2, n = 10; M 4 /Ca V 1.2, n = 5). Statistical significance was evaluated with an unpaired Student’s t test.

    Article Snippet: First, it was incubated with one of the following primary antibodies (Abs): rabbit anti-caveolin-1 (sc-894: Santa Cruz Biotechnology, Santa Cruz, CA, USA) (RRID:AB_2072042), mouse anti-transferrin receptor (A11130: Molecular Probes, Eugene, OR, USA) (RRID:AB_2534136), mouse anti-M 4 (MAB1576: Chemicon, Temecula, CA, USA) (RRID:AB_2080217), rabbit anti-Ca V 1.2 (ACC-001: Alomone, Jerusalem, Israel) (RRID:AB_2039764), or rabbit anti-TWIK-related acid-sensitive K + 1 (TASK1) (APC-024: Alomone) (RRID:AB_2040132).

    Techniques: Membrane, Immunostaining, Transfection, Construct, Labeling, Immunofluorescence, Fluorescence, Fractionation, Gradient Centrifugation, Marker, Double Staining

    Diagram showing localization of caveolin-1, Ca V 1.2, muscarinic M 4 receptor subtype, and TASK1 in the raft and non-raft membrane domains.

    Journal: Acta Histochemica et Cytochemica

    Article Title: Muscarinic Receptor Stimulation Does Not Inhibit Voltage-dependent Ca 2+ Channels in Rat Adrenal Medullary Chromaffin Cells

    doi: 10.1267/ahc.23-00042

    Figure Lengend Snippet: Diagram showing localization of caveolin-1, Ca V 1.2, muscarinic M 4 receptor subtype, and TASK1 in the raft and non-raft membrane domains.

    Article Snippet: First, it was incubated with one of the following primary antibodies (Abs): rabbit anti-caveolin-1 (sc-894: Santa Cruz Biotechnology, Santa Cruz, CA, USA) (RRID:AB_2072042), mouse anti-transferrin receptor (A11130: Molecular Probes, Eugene, OR, USA) (RRID:AB_2534136), mouse anti-M 4 (MAB1576: Chemicon, Temecula, CA, USA) (RRID:AB_2080217), rabbit anti-Ca V 1.2 (ACC-001: Alomone, Jerusalem, Israel) (RRID:AB_2039764), or rabbit anti-TWIK-related acid-sensitive K + 1 (TASK1) (APC-024: Alomone) (RRID:AB_2040132).

    Techniques: Membrane