anti cav1 2  (Bioss)


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

    Structured Review

    Bioss anti cav1 2
    Anti Cav1 2, supplied by Bioss, 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/anti cav1 2/product/Bioss
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cav1 2 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    cav1 2 protein quantificationwas determinedwith bio rad image lab software  (Bio-Rad)

     
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Bio-Rad cav1 2 protein quantificationwas determinedwith bio rad image lab software
    Cav1 2 Protein Quantificationwas Determinedwith Bio Rad Image Lab Software, supplied by Bio-Rad, 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/cav1 2 protein quantificationwas determinedwith bio rad image lab software/product/Bio-Rad
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cav1 2 protein quantificationwas determinedwith bio rad image lab software - by Bioz Stars, 2024-07
    86/100 stars

    Images

    cav1 2  (Alomone Labs)


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

    Structured Review

    Alomone Labs cav1 2
    Cav1 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/cav1 2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cav1 2 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    anti cav1 2  (Alomone Labs)


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

    Structured Review

    Alomone Labs anti cav1 2
    Anti Cav1 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/anti cav1 2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cav1 2 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    cav1 2  (Thermo Fisher)


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

    Structured Review

    Thermo Fisher cav1 2
    Cav1 2, supplied by Thermo Fisher, 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/cav1 2/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cav1 2 - by Bioz Stars, 2024-07
    86/100 stars

    Images


    Structured Review

    Dawley Inc ⬇ cav1 2 channel protein trafficking
    The mechanisms of allicin in arrhythmias
    ⬇ Cav1 2 Channel Protein Trafficking, supplied by Dawley Inc, 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/⬇ cav1 2 channel protein trafficking/product/Dawley Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ⬇ cav1 2 channel protein trafficking - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "Therapeutic potentials of allicin in cardiovascular disease: advances and future directions"

    Article Title: Therapeutic potentials of allicin in cardiovascular disease: advances and future directions

    Journal: Chinese Medicine

    doi: 10.1186/s13020-024-00936-8

    The mechanisms of allicin in arrhythmias
    Figure Legend Snippet: The mechanisms of allicin in arrhythmias

    Techniques Used: Concentration Assay, Plasmid Preparation


    Structured Review

    Millipore anti cav1 2 antibody
    a , SEM images of vertical nanopillars. Scale bar 1 µm on the left, 500 nm on the right. b , Schematics of the T-tubule system in a cardiomyocyte (left), ER-PM contacts formed at T-tubules (middle), and nanopillar-induced membrane invaginations (right). c-d , Co-Immunostained JPH2 (green) and α-actinin (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on flat areas (c) and on nanopillar areas (d). JPH2 (green) preferentially accumulates on nanopillar locations. Magnified images of the yellow boxes are shown at the bottom. Scale bar 10 µm on the top, 2.5 µm at the bottom. e , Averaged fluorescent signals for GFP-Sec61β, JPH2, and α-actinin. 101×101-pixel square regions of interest centered by each nanopillar were averaged and displayed. Over ∼3000 nanopillars were included in each condition. GFP-Sec61β n = 18 cells; JPH2 and α-actinin n = 21 cells. f , Intensity plots along the horizontal yellow line in e, normalized by the intensity value of the region between two nanopillars. Error bars represent standard deviation (STD) among values calculated from individual cells. g , The ratio of the fluorescent intensity at the nanopillars (pillar: area in the magenta mask) over the average intensity between the nanopillars (non-pillar: area inside the yellow mask). n=21, 10, 11, 18, 21 cells for JPH2, RyR2, <t>Cav1.2,</t> Sec61β, and α-actinin. ****P <0.0001; **P=0.0067; not-significant (NS) P>0.9999. h , Representative images of co-immunostained RyR2 (green) and Cav1.2 (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on nanopillars. The solid arrowhead points at a cell with clear Cav1.2 signal on nanopillars and the hollow arrowhead points to a cell with minimal Cav1.2 on nanopillars. Scale bar 10 µm on the top, 2.5 µm on the bottom. i , Averaged fluorescent signals of ∼1200 nanopillars were included in each condition. n = 10 cells for RyR2; n = 11 cells for Cav1.2. j , SEM images of nanobars. Scale bar 5 µm on the top, 2 µm at the bottom. k , Representative images of iPSC-CMs with expressed BFP-CAAX, immunostaining of JPH2, RyR2, and Cav1.2 on nanobars. The RyR2 staining and Cav1.2 staining are the same cell. Magnified images of the yellow boxes are shown in the middle row. Averaged fluorescent signals of all nanobars from multiple cells were averaged and displayed in the bottom row. n = 23, 23, 23, 11 cells for BFP-CAAX, JPH2, RyR2, and Cav1.2. Scale bar 10 µm in the top row, 2.5 µm in the middle row. l , Quantification of the ratio of the fluorescent intensity at the ends of nanobars (area in the magenta mask) over the average at the sides of nanobars (area in the green mask). Cell number is the same as in ( k ). ****P <0.0001; *P=0.0277. All experiments were independently replicated at least three times. All error bars represent STD. Kruskal-Wallis test corrected with Dunn’s multiple-comparison test was used to assess significance for ( g , l ).
    Anti Cav1 2 Antibody, supplied by Millipore, 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/anti cav1 2 antibody/product/Millipore
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cav1 2 antibody - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "Membrane Curvature Promotes ER-PM Contact Formation via Junctophilin-EHD Interactions"

    Article Title: Membrane Curvature Promotes ER-PM Contact Formation via Junctophilin-EHD Interactions

    Journal: bioRxiv

    doi: 10.1101/2024.06.29.601287

    a , SEM images of vertical nanopillars. Scale bar 1 µm on the left, 500 nm on the right. b , Schematics of the T-tubule system in a cardiomyocyte (left), ER-PM contacts formed at T-tubules (middle), and nanopillar-induced membrane invaginations (right). c-d , Co-Immunostained JPH2 (green) and α-actinin (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on flat areas (c) and on nanopillar areas (d). JPH2 (green) preferentially accumulates on nanopillar locations. Magnified images of the yellow boxes are shown at the bottom. Scale bar 10 µm on the top, 2.5 µm at the bottom. e , Averaged fluorescent signals for GFP-Sec61β, JPH2, and α-actinin. 101×101-pixel square regions of interest centered by each nanopillar were averaged and displayed. Over ∼3000 nanopillars were included in each condition. GFP-Sec61β n = 18 cells; JPH2 and α-actinin n = 21 cells. f , Intensity plots along the horizontal yellow line in e, normalized by the intensity value of the region between two nanopillars. Error bars represent standard deviation (STD) among values calculated from individual cells. g , The ratio of the fluorescent intensity at the nanopillars (pillar: area in the magenta mask) over the average intensity between the nanopillars (non-pillar: area inside the yellow mask). n=21, 10, 11, 18, 21 cells for JPH2, RyR2, Cav1.2, Sec61β, and α-actinin. ****P <0.0001; **P=0.0067; not-significant (NS) P>0.9999. h , Representative images of co-immunostained RyR2 (green) and Cav1.2 (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on nanopillars. The solid arrowhead points at a cell with clear Cav1.2 signal on nanopillars and the hollow arrowhead points to a cell with minimal Cav1.2 on nanopillars. Scale bar 10 µm on the top, 2.5 µm on the bottom. i , Averaged fluorescent signals of ∼1200 nanopillars were included in each condition. n = 10 cells for RyR2; n = 11 cells for Cav1.2. j , SEM images of nanobars. Scale bar 5 µm on the top, 2 µm at the bottom. k , Representative images of iPSC-CMs with expressed BFP-CAAX, immunostaining of JPH2, RyR2, and Cav1.2 on nanobars. The RyR2 staining and Cav1.2 staining are the same cell. Magnified images of the yellow boxes are shown in the middle row. Averaged fluorescent signals of all nanobars from multiple cells were averaged and displayed in the bottom row. n = 23, 23, 23, 11 cells for BFP-CAAX, JPH2, RyR2, and Cav1.2. Scale bar 10 µm in the top row, 2.5 µm in the middle row. l , Quantification of the ratio of the fluorescent intensity at the ends of nanobars (area in the magenta mask) over the average at the sides of nanobars (area in the green mask). Cell number is the same as in ( k ). ****P <0.0001; *P=0.0277. All experiments were independently replicated at least three times. All error bars represent STD. Kruskal-Wallis test corrected with Dunn’s multiple-comparison test was used to assess significance for ( g , l ).
    Figure Legend Snippet: a , SEM images of vertical nanopillars. Scale bar 1 µm on the left, 500 nm on the right. b , Schematics of the T-tubule system in a cardiomyocyte (left), ER-PM contacts formed at T-tubules (middle), and nanopillar-induced membrane invaginations (right). c-d , Co-Immunostained JPH2 (green) and α-actinin (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on flat areas (c) and on nanopillar areas (d). JPH2 (green) preferentially accumulates on nanopillar locations. Magnified images of the yellow boxes are shown at the bottom. Scale bar 10 µm on the top, 2.5 µm at the bottom. e , Averaged fluorescent signals for GFP-Sec61β, JPH2, and α-actinin. 101×101-pixel square regions of interest centered by each nanopillar were averaged and displayed. Over ∼3000 nanopillars were included in each condition. GFP-Sec61β n = 18 cells; JPH2 and α-actinin n = 21 cells. f , Intensity plots along the horizontal yellow line in e, normalized by the intensity value of the region between two nanopillars. Error bars represent standard deviation (STD) among values calculated from individual cells. g , The ratio of the fluorescent intensity at the nanopillars (pillar: area in the magenta mask) over the average intensity between the nanopillars (non-pillar: area inside the yellow mask). n=21, 10, 11, 18, 21 cells for JPH2, RyR2, Cav1.2, Sec61β, and α-actinin. ****P <0.0001; **P=0.0067; not-significant (NS) P>0.9999. h , Representative images of co-immunostained RyR2 (green) and Cav1.2 (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on nanopillars. The solid arrowhead points at a cell with clear Cav1.2 signal on nanopillars and the hollow arrowhead points to a cell with minimal Cav1.2 on nanopillars. Scale bar 10 µm on the top, 2.5 µm on the bottom. i , Averaged fluorescent signals of ∼1200 nanopillars were included in each condition. n = 10 cells for RyR2; n = 11 cells for Cav1.2. j , SEM images of nanobars. Scale bar 5 µm on the top, 2 µm at the bottom. k , Representative images of iPSC-CMs with expressed BFP-CAAX, immunostaining of JPH2, RyR2, and Cav1.2 on nanobars. The RyR2 staining and Cav1.2 staining are the same cell. Magnified images of the yellow boxes are shown in the middle row. Averaged fluorescent signals of all nanobars from multiple cells were averaged and displayed in the bottom row. n = 23, 23, 23, 11 cells for BFP-CAAX, JPH2, RyR2, and Cav1.2. Scale bar 10 µm in the top row, 2.5 µm in the middle row. l , Quantification of the ratio of the fluorescent intensity at the ends of nanobars (area in the magenta mask) over the average at the sides of nanobars (area in the green mask). Cell number is the same as in ( k ). ****P <0.0001; *P=0.0277. All experiments were independently replicated at least three times. All error bars represent STD. Kruskal-Wallis test corrected with Dunn’s multiple-comparison test was used to assess significance for ( g , l ).

    Techniques Used: Membrane, Expressing, Standard Deviation, Immunostaining, Staining, Comparison

    a , An illustration of the overlapping between the PM targeting and the curvature sensing candidates in JPH2 interactome. b , Representative images of GFP-JPH3 on nanobars shRNA knockdown of scramble, Clathrin, CAVIN1, CAV1/2, or EHD1/2/4. ShRNAs sequences were fused with fluorescence proteins to identify knockdown cells. Bright field and the fluorescent protein expression are shown on the left. Zooms are the enlarged view of the region in yellow boxes. Averaged nanobar images from multiple cells are shown on the right. Cell number for average: scramble = 28, shCLTC (Clathrin heavy chain) = 23, shCAV1/2 = 53, shCAVIN1 = 30, shEHD1/2/4 = 55. Scale bar 10 µm in whole cell images, 5 µm in zoom-in images. c , Quantifications of GFP-JPH3 end-to-side ratios upon shRNA knockdowns. Conditions and cell numbers are the same as in b. ****P<0.0001, NS(shClathrin) P>0.9999, NS(shCAV1shCAV2) P>0.9999, NS(shCAVIN1) P=0.0592. d , The distribution of GFP-JPH3 on nanobars before and after 10 mM MβCD treatment at 37°C for 30 min. Scale bar 5 µm. Right: averaged images of all bars in a single cell. e , Quantifications of the nanobar end-to-side ratios for GFP-JPH3 in U2OS cells before and after the MβCD treatment. n = 15 cells for each group. f , Representative images of U2OS cells co-expressing EHD4-GFP/mCherry with indicated JPH3 mutants. Enlarged views are zoomed from the yellow boxes in whole cell image on the left. Scale bar 10 µm in whole cell images, 5 µm in zoom-in images. g , Pearson’s correlation coefficients (PCC) between coexpressed EHD4-GFP/mCherry and JPH3 mutants shown in ( f ). Cell number: mCherry-8MORN_LCR = 21, GFP-8MORN_LCR-αHelix = 17, mCherry-LCR = 19, GFP-8MORN-αHelix = 19. ****P < 0.0001, **P = 0.0024 (8MORN_LCR-αHlx vs. 8MORN-αHlx), **P = 0.0075 (8MORN_LCR vs. 8MORN-αHlx), *P = 0.0174 (8MORN-αHlx vs. LCR), NS P= 0.9995 (8MORN_LCR-αHlx vs. 8MORN_LCR). h , PCC for colocalization between mCherry-8MORN_LCR and EHD4-GFP or between mCherry-8MORN_LCR and anti-clathrin. n = 21 cells for each group. ****P<0.0001. i , Co-immunoprecipitation (IP) from HEK cells expressing EHD4-GFP and mCherry-8MORN_LCR or mCherry-LCR. GFP was pulled via GFP-antibody beads, and the immunoprecipitates were blotted with mCherry antibody. mCherry-8MORN_LCR, but not mCherry-LCR, was pulled together with EHD4-GFP. All experiments were independently replicated at least three times. All error bars represent STD. Kruskal–Wallis test corrected with Dunn’s multiple-comparison test was used for ( c ). Unpaired two-tailed Welch’s t test was used to assess significance for ( e , h ). Brown-Forsythe and Welch ANOVA tests was used to assess significance for ( g ).
    Figure Legend Snippet: a , An illustration of the overlapping between the PM targeting and the curvature sensing candidates in JPH2 interactome. b , Representative images of GFP-JPH3 on nanobars shRNA knockdown of scramble, Clathrin, CAVIN1, CAV1/2, or EHD1/2/4. ShRNAs sequences were fused with fluorescence proteins to identify knockdown cells. Bright field and the fluorescent protein expression are shown on the left. Zooms are the enlarged view of the region in yellow boxes. Averaged nanobar images from multiple cells are shown on the right. Cell number for average: scramble = 28, shCLTC (Clathrin heavy chain) = 23, shCAV1/2 = 53, shCAVIN1 = 30, shEHD1/2/4 = 55. Scale bar 10 µm in whole cell images, 5 µm in zoom-in images. c , Quantifications of GFP-JPH3 end-to-side ratios upon shRNA knockdowns. Conditions and cell numbers are the same as in b. ****P<0.0001, NS(shClathrin) P>0.9999, NS(shCAV1shCAV2) P>0.9999, NS(shCAVIN1) P=0.0592. d , The distribution of GFP-JPH3 on nanobars before and after 10 mM MβCD treatment at 37°C for 30 min. Scale bar 5 µm. Right: averaged images of all bars in a single cell. e , Quantifications of the nanobar end-to-side ratios for GFP-JPH3 in U2OS cells before and after the MβCD treatment. n = 15 cells for each group. f , Representative images of U2OS cells co-expressing EHD4-GFP/mCherry with indicated JPH3 mutants. Enlarged views are zoomed from the yellow boxes in whole cell image on the left. Scale bar 10 µm in whole cell images, 5 µm in zoom-in images. g , Pearson’s correlation coefficients (PCC) between coexpressed EHD4-GFP/mCherry and JPH3 mutants shown in ( f ). Cell number: mCherry-8MORN_LCR = 21, GFP-8MORN_LCR-αHelix = 17, mCherry-LCR = 19, GFP-8MORN-αHelix = 19. ****P < 0.0001, **P = 0.0024 (8MORN_LCR-αHlx vs. 8MORN-αHlx), **P = 0.0075 (8MORN_LCR vs. 8MORN-αHlx), *P = 0.0174 (8MORN-αHlx vs. LCR), NS P= 0.9995 (8MORN_LCR-αHlx vs. 8MORN_LCR). h , PCC for colocalization between mCherry-8MORN_LCR and EHD4-GFP or between mCherry-8MORN_LCR and anti-clathrin. n = 21 cells for each group. ****P<0.0001. i , Co-immunoprecipitation (IP) from HEK cells expressing EHD4-GFP and mCherry-8MORN_LCR or mCherry-LCR. GFP was pulled via GFP-antibody beads, and the immunoprecipitates were blotted with mCherry antibody. mCherry-8MORN_LCR, but not mCherry-LCR, was pulled together with EHD4-GFP. All experiments were independently replicated at least three times. All error bars represent STD. Kruskal–Wallis test corrected with Dunn’s multiple-comparison test was used for ( c ). Unpaired two-tailed Welch’s t test was used to assess significance for ( e , h ). Brown-Forsythe and Welch ANOVA tests was used to assess significance for ( g ).

    Techniques Used: shRNA, Knockdown, Fluorescence, Expressing, Immunoprecipitation, Comparison, Two Tailed Test

    a , EHD4, CAVIN1, Caveolin1, and Caveolin2 all show preferential accumulations at the nanobar ends. Quantifications of the end-to-side ratios are shown on the right. Cell number: n =14 cells for Caveolin2, n = 15 cells for all other groups. ***P=0.0001, **P=0.0015; ****P<0.0001. b , mRNA transcript levels in U2OS cells (shown as normalized transcripts per million, nTPM), obtained from protein atlas database. Genes highlighted in gray: not identified in the JPH2 interactome but are closely related either genetically or functionally. Genes including EHD3, CAVIN2, CAVIN3, and CAVIN4 were not selected as knockdown targets in KD experiments because of their low TPM level in U2OS cells. c , Western blots confirming the effective shRNA knockdown of CAV1, CAV2, EHD1, EHD2, CLTC (clathrin heavy chain), and BIN1. GAPDH was blotted as a loading control. d , Representative images of U2OS cells co-expressing EHD4-mCherry with EHD1-GFP (Top) or with EHD2-GFP (Bottom). Enlarged views are zoomed from the yellow boxes in the whole cell images. e , Representative images of U2OS cells expressing GFP-JPH3 (green) and EHD4-mCh (magenta) seeding on nanobars. Enlarged views are zoomed from the yellow boxes in whole cell images. f , Representative images of immunostaining of EHD2 or EHD4 in iPSC-CMs or rat embryonic CMs on nanopillars. Magnified images of the region in yellow boxes are shown in the bottom row. Scale bar 10 µm in the top row, 2.5 µm in the bottom row. g , Representative images of immunostaining of EHD4 in iPSC-CMs on nanobars. Magnified images of the region in yellow boxes are shown in the middle row. Averaged fluorescent signals of all nanobars from multiple cells were averaged and displayed in the bottom row. n = 16 cells for the average image. Bottom graph: quantification of the end to side ratios of EHD4 compared to that of BFP-CAAX expressed in iPSC-CM on nanobars. Cell number: CAAX = 23 (same data as in ), EHD4 = 16. ****P <0.0001. h , Representative images of immunostaining of JPH2 in iPSC cells transfected with shRNA of scramble (top) or EHD1/2/4 (bottom). From left to right: Bright field image, fluorescent protein expression indicating successful transfection of shRNA, and JPH2 staining. Averaged nanobar images of JPH2 staining from multiple cells are shown at the bottom of JPH staining panels. Quantifications of the end-to-side ratios are shown on the right. Cell number for both average and quantification: Scramble = 24, shEHD1/2/4 = 26. ****P <0.0001. Scale bars 10 µm in all whole cell images, 5 µm in all zoomed images unless otherwise mentioned. All experiments were independently replicated at least two times. All error bars represent STD. Kruskal–Wallis test corrected with Dunn’s multiple-comparison test was used to assess significance in ( a ). Unpaired two-tailed Welch’s t test was used to assess significance in ( g ). Two-tailed Mann-Whitney test was used to assess significance in ( h ).
    Figure Legend Snippet: a , EHD4, CAVIN1, Caveolin1, and Caveolin2 all show preferential accumulations at the nanobar ends. Quantifications of the end-to-side ratios are shown on the right. Cell number: n =14 cells for Caveolin2, n = 15 cells for all other groups. ***P=0.0001, **P=0.0015; ****P<0.0001. b , mRNA transcript levels in U2OS cells (shown as normalized transcripts per million, nTPM), obtained from protein atlas database. Genes highlighted in gray: not identified in the JPH2 interactome but are closely related either genetically or functionally. Genes including EHD3, CAVIN2, CAVIN3, and CAVIN4 were not selected as knockdown targets in KD experiments because of their low TPM level in U2OS cells. c , Western blots confirming the effective shRNA knockdown of CAV1, CAV2, EHD1, EHD2, CLTC (clathrin heavy chain), and BIN1. GAPDH was blotted as a loading control. d , Representative images of U2OS cells co-expressing EHD4-mCherry with EHD1-GFP (Top) or with EHD2-GFP (Bottom). Enlarged views are zoomed from the yellow boxes in the whole cell images. e , Representative images of U2OS cells expressing GFP-JPH3 (green) and EHD4-mCh (magenta) seeding on nanobars. Enlarged views are zoomed from the yellow boxes in whole cell images. f , Representative images of immunostaining of EHD2 or EHD4 in iPSC-CMs or rat embryonic CMs on nanopillars. Magnified images of the region in yellow boxes are shown in the bottom row. Scale bar 10 µm in the top row, 2.5 µm in the bottom row. g , Representative images of immunostaining of EHD4 in iPSC-CMs on nanobars. Magnified images of the region in yellow boxes are shown in the middle row. Averaged fluorescent signals of all nanobars from multiple cells were averaged and displayed in the bottom row. n = 16 cells for the average image. Bottom graph: quantification of the end to side ratios of EHD4 compared to that of BFP-CAAX expressed in iPSC-CM on nanobars. Cell number: CAAX = 23 (same data as in ), EHD4 = 16. ****P <0.0001. h , Representative images of immunostaining of JPH2 in iPSC cells transfected with shRNA of scramble (top) or EHD1/2/4 (bottom). From left to right: Bright field image, fluorescent protein expression indicating successful transfection of shRNA, and JPH2 staining. Averaged nanobar images of JPH2 staining from multiple cells are shown at the bottom of JPH staining panels. Quantifications of the end-to-side ratios are shown on the right. Cell number for both average and quantification: Scramble = 24, shEHD1/2/4 = 26. ****P <0.0001. Scale bars 10 µm in all whole cell images, 5 µm in all zoomed images unless otherwise mentioned. All experiments were independently replicated at least two times. All error bars represent STD. Kruskal–Wallis test corrected with Dunn’s multiple-comparison test was used to assess significance in ( a ). Unpaired two-tailed Welch’s t test was used to assess significance in ( g ). Two-tailed Mann-Whitney test was used to assess significance in ( h ).

    Techniques Used: Knockdown, Western Blot, shRNA, Control, Expressing, Immunostaining, Transfection, Staining, Comparison, Two Tailed Test, MANN-WHITNEY

    anti cav1 2 cacna1c antibody  (Thermo Fisher)


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

    Structured Review

    Thermo Fisher anti cav1 2 cacna1c antibody
    Anti Cav1 2 Cacna1c Antibody, supplied by Thermo Fisher, 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/anti cav1 2 cacna1c antibody/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cav1 2 cacna1c antibody - by Bioz Stars, 2024-07
    86/100 stars

    Images


    Structured Review

    NeuroMab ltcc cav1 2 neuromab clone n263 31
    Ltcc Cav1 2 Neuromab Clone N263 31, supplied by NeuroMab, 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/ltcc cav1 2 neuromab clone n263 31/product/NeuroMab
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ltcc cav1 2 neuromab clone n263 31 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    cav1 2  (Alomone Labs)


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

    Structured Review

    Alomone Labs cav1 2
    Cav1 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/cav1 2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cav1 2 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86
    Bioss anti cav1 2
    Anti Cav1 2, supplied by Bioss, 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/anti cav1 2/product/Bioss
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cav1 2 - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Bio-Rad cav1 2 protein quantificationwas determinedwith bio rad image lab software
    Cav1 2 Protein Quantificationwas Determinedwith Bio Rad Image Lab Software, supplied by Bio-Rad, 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/cav1 2 protein quantificationwas determinedwith bio rad image lab software/product/Bio-Rad
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cav1 2 protein quantificationwas determinedwith bio rad image lab software - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Alomone Labs cav1 2
    Cav1 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/cav1 2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cav1 2 - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Alomone Labs anti cav1 2
    Anti Cav1 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/anti cav1 2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cav1 2 - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Thermo Fisher cav1 2
    Cav1 2, supplied by Thermo Fisher, 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/cav1 2/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cav1 2 - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Dawley Inc ⬇ cav1 2 channel protein trafficking
    The mechanisms of allicin in arrhythmias
    ⬇ Cav1 2 Channel Protein Trafficking, supplied by Dawley Inc, 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/⬇ cav1 2 channel protein trafficking/product/Dawley Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ⬇ cav1 2 channel protein trafficking - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Millipore anti cav1 2 antibody
    a , SEM images of vertical nanopillars. Scale bar 1 µm on the left, 500 nm on the right. b , Schematics of the T-tubule system in a cardiomyocyte (left), ER-PM contacts formed at T-tubules (middle), and nanopillar-induced membrane invaginations (right). c-d , Co-Immunostained JPH2 (green) and α-actinin (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on flat areas (c) and on nanopillar areas (d). JPH2 (green) preferentially accumulates on nanopillar locations. Magnified images of the yellow boxes are shown at the bottom. Scale bar 10 µm on the top, 2.5 µm at the bottom. e , Averaged fluorescent signals for GFP-Sec61β, JPH2, and α-actinin. 101×101-pixel square regions of interest centered by each nanopillar were averaged and displayed. Over ∼3000 nanopillars were included in each condition. GFP-Sec61β n = 18 cells; JPH2 and α-actinin n = 21 cells. f , Intensity plots along the horizontal yellow line in e, normalized by the intensity value of the region between two nanopillars. Error bars represent standard deviation (STD) among values calculated from individual cells. g , The ratio of the fluorescent intensity at the nanopillars (pillar: area in the magenta mask) over the average intensity between the nanopillars (non-pillar: area inside the yellow mask). n=21, 10, 11, 18, 21 cells for JPH2, RyR2, <t>Cav1.2,</t> Sec61β, and α-actinin. ****P <0.0001; **P=0.0067; not-significant (NS) P>0.9999. h , Representative images of co-immunostained RyR2 (green) and Cav1.2 (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on nanopillars. The solid arrowhead points at a cell with clear Cav1.2 signal on nanopillars and the hollow arrowhead points to a cell with minimal Cav1.2 on nanopillars. Scale bar 10 µm on the top, 2.5 µm on the bottom. i , Averaged fluorescent signals of ∼1200 nanopillars were included in each condition. n = 10 cells for RyR2; n = 11 cells for Cav1.2. j , SEM images of nanobars. Scale bar 5 µm on the top, 2 µm at the bottom. k , Representative images of iPSC-CMs with expressed BFP-CAAX, immunostaining of JPH2, RyR2, and Cav1.2 on nanobars. The RyR2 staining and Cav1.2 staining are the same cell. Magnified images of the yellow boxes are shown in the middle row. Averaged fluorescent signals of all nanobars from multiple cells were averaged and displayed in the bottom row. n = 23, 23, 23, 11 cells for BFP-CAAX, JPH2, RyR2, and Cav1.2. Scale bar 10 µm in the top row, 2.5 µm in the middle row. l , Quantification of the ratio of the fluorescent intensity at the ends of nanobars (area in the magenta mask) over the average at the sides of nanobars (area in the green mask). Cell number is the same as in ( k ). ****P <0.0001; *P=0.0277. All experiments were independently replicated at least three times. All error bars represent STD. Kruskal-Wallis test corrected with Dunn’s multiple-comparison test was used to assess significance for ( g , l ).
    Anti Cav1 2 Antibody, supplied by Millipore, 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/anti cav1 2 antibody/product/Millipore
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cav1 2 antibody - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Thermo Fisher anti cav1 2 cacna1c antibody
    a , SEM images of vertical nanopillars. Scale bar 1 µm on the left, 500 nm on the right. b , Schematics of the T-tubule system in a cardiomyocyte (left), ER-PM contacts formed at T-tubules (middle), and nanopillar-induced membrane invaginations (right). c-d , Co-Immunostained JPH2 (green) and α-actinin (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on flat areas (c) and on nanopillar areas (d). JPH2 (green) preferentially accumulates on nanopillar locations. Magnified images of the yellow boxes are shown at the bottom. Scale bar 10 µm on the top, 2.5 µm at the bottom. e , Averaged fluorescent signals for GFP-Sec61β, JPH2, and α-actinin. 101×101-pixel square regions of interest centered by each nanopillar were averaged and displayed. Over ∼3000 nanopillars were included in each condition. GFP-Sec61β n = 18 cells; JPH2 and α-actinin n = 21 cells. f , Intensity plots along the horizontal yellow line in e, normalized by the intensity value of the region between two nanopillars. Error bars represent standard deviation (STD) among values calculated from individual cells. g , The ratio of the fluorescent intensity at the nanopillars (pillar: area in the magenta mask) over the average intensity between the nanopillars (non-pillar: area inside the yellow mask). n=21, 10, 11, 18, 21 cells for JPH2, RyR2, <t>Cav1.2,</t> Sec61β, and α-actinin. ****P <0.0001; **P=0.0067; not-significant (NS) P>0.9999. h , Representative images of co-immunostained RyR2 (green) and Cav1.2 (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on nanopillars. The solid arrowhead points at a cell with clear Cav1.2 signal on nanopillars and the hollow arrowhead points to a cell with minimal Cav1.2 on nanopillars. Scale bar 10 µm on the top, 2.5 µm on the bottom. i , Averaged fluorescent signals of ∼1200 nanopillars were included in each condition. n = 10 cells for RyR2; n = 11 cells for Cav1.2. j , SEM images of nanobars. Scale bar 5 µm on the top, 2 µm at the bottom. k , Representative images of iPSC-CMs with expressed BFP-CAAX, immunostaining of JPH2, RyR2, and Cav1.2 on nanobars. The RyR2 staining and Cav1.2 staining are the same cell. Magnified images of the yellow boxes are shown in the middle row. Averaged fluorescent signals of all nanobars from multiple cells were averaged and displayed in the bottom row. n = 23, 23, 23, 11 cells for BFP-CAAX, JPH2, RyR2, and Cav1.2. Scale bar 10 µm in the top row, 2.5 µm in the middle row. l , Quantification of the ratio of the fluorescent intensity at the ends of nanobars (area in the magenta mask) over the average at the sides of nanobars (area in the green mask). Cell number is the same as in ( k ). ****P <0.0001; *P=0.0277. All experiments were independently replicated at least three times. All error bars represent STD. Kruskal-Wallis test corrected with Dunn’s multiple-comparison test was used to assess significance for ( g , l ).
    Anti Cav1 2 Cacna1c Antibody, supplied by Thermo Fisher, 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/anti cav1 2 cacna1c antibody/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cav1 2 cacna1c antibody - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    NeuroMab ltcc cav1 2 neuromab clone n263 31
    a , SEM images of vertical nanopillars. Scale bar 1 µm on the left, 500 nm on the right. b , Schematics of the T-tubule system in a cardiomyocyte (left), ER-PM contacts formed at T-tubules (middle), and nanopillar-induced membrane invaginations (right). c-d , Co-Immunostained JPH2 (green) and α-actinin (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on flat areas (c) and on nanopillar areas (d). JPH2 (green) preferentially accumulates on nanopillar locations. Magnified images of the yellow boxes are shown at the bottom. Scale bar 10 µm on the top, 2.5 µm at the bottom. e , Averaged fluorescent signals for GFP-Sec61β, JPH2, and α-actinin. 101×101-pixel square regions of interest centered by each nanopillar were averaged and displayed. Over ∼3000 nanopillars were included in each condition. GFP-Sec61β n = 18 cells; JPH2 and α-actinin n = 21 cells. f , Intensity plots along the horizontal yellow line in e, normalized by the intensity value of the region between two nanopillars. Error bars represent standard deviation (STD) among values calculated from individual cells. g , The ratio of the fluorescent intensity at the nanopillars (pillar: area in the magenta mask) over the average intensity between the nanopillars (non-pillar: area inside the yellow mask). n=21, 10, 11, 18, 21 cells for JPH2, RyR2, <t>Cav1.2,</t> Sec61β, and α-actinin. ****P <0.0001; **P=0.0067; not-significant (NS) P>0.9999. h , Representative images of co-immunostained RyR2 (green) and Cav1.2 (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on nanopillars. The solid arrowhead points at a cell with clear Cav1.2 signal on nanopillars and the hollow arrowhead points to a cell with minimal Cav1.2 on nanopillars. Scale bar 10 µm on the top, 2.5 µm on the bottom. i , Averaged fluorescent signals of ∼1200 nanopillars were included in each condition. n = 10 cells for RyR2; n = 11 cells for Cav1.2. j , SEM images of nanobars. Scale bar 5 µm on the top, 2 µm at the bottom. k , Representative images of iPSC-CMs with expressed BFP-CAAX, immunostaining of JPH2, RyR2, and Cav1.2 on nanobars. The RyR2 staining and Cav1.2 staining are the same cell. Magnified images of the yellow boxes are shown in the middle row. Averaged fluorescent signals of all nanobars from multiple cells were averaged and displayed in the bottom row. n = 23, 23, 23, 11 cells for BFP-CAAX, JPH2, RyR2, and Cav1.2. Scale bar 10 µm in the top row, 2.5 µm in the middle row. l , Quantification of the ratio of the fluorescent intensity at the ends of nanobars (area in the magenta mask) over the average at the sides of nanobars (area in the green mask). Cell number is the same as in ( k ). ****P <0.0001; *P=0.0277. All experiments were independently replicated at least three times. All error bars represent STD. Kruskal-Wallis test corrected with Dunn’s multiple-comparison test was used to assess significance for ( g , l ).
    Ltcc Cav1 2 Neuromab Clone N263 31, supplied by NeuroMab, 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/ltcc cav1 2 neuromab clone n263 31/product/NeuroMab
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ltcc cav1 2 neuromab clone n263 31 - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    Image Search Results


    The mechanisms of allicin in arrhythmias

    Journal: Chinese Medicine

    Article Title: Therapeutic potentials of allicin in cardiovascular disease: advances and future directions

    doi: 10.1186/s13020-024-00936-8

    Figure Lengend Snippet: The mechanisms of allicin in arrhythmias

    Article Snippet: Primary cardiomyocytes from neonatal Sprague–Dawley rats , – , 3, 10, 30 μmol/l for 48 h , Anti-arrhythmia , ⬇ Cav1.2 channel protein trafficking , [ ] .

    Techniques: Concentration Assay, Plasmid Preparation

    a , SEM images of vertical nanopillars. Scale bar 1 µm on the left, 500 nm on the right. b , Schematics of the T-tubule system in a cardiomyocyte (left), ER-PM contacts formed at T-tubules (middle), and nanopillar-induced membrane invaginations (right). c-d , Co-Immunostained JPH2 (green) and α-actinin (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on flat areas (c) and on nanopillar areas (d). JPH2 (green) preferentially accumulates on nanopillar locations. Magnified images of the yellow boxes are shown at the bottom. Scale bar 10 µm on the top, 2.5 µm at the bottom. e , Averaged fluorescent signals for GFP-Sec61β, JPH2, and α-actinin. 101×101-pixel square regions of interest centered by each nanopillar were averaged and displayed. Over ∼3000 nanopillars were included in each condition. GFP-Sec61β n = 18 cells; JPH2 and α-actinin n = 21 cells. f , Intensity plots along the horizontal yellow line in e, normalized by the intensity value of the region between two nanopillars. Error bars represent standard deviation (STD) among values calculated from individual cells. g , The ratio of the fluorescent intensity at the nanopillars (pillar: area in the magenta mask) over the average intensity between the nanopillars (non-pillar: area inside the yellow mask). n=21, 10, 11, 18, 21 cells for JPH2, RyR2, Cav1.2, Sec61β, and α-actinin. ****P <0.0001; **P=0.0067; not-significant (NS) P>0.9999. h , Representative images of co-immunostained RyR2 (green) and Cav1.2 (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on nanopillars. The solid arrowhead points at a cell with clear Cav1.2 signal on nanopillars and the hollow arrowhead points to a cell with minimal Cav1.2 on nanopillars. Scale bar 10 µm on the top, 2.5 µm on the bottom. i , Averaged fluorescent signals of ∼1200 nanopillars were included in each condition. n = 10 cells for RyR2; n = 11 cells for Cav1.2. j , SEM images of nanobars. Scale bar 5 µm on the top, 2 µm at the bottom. k , Representative images of iPSC-CMs with expressed BFP-CAAX, immunostaining of JPH2, RyR2, and Cav1.2 on nanobars. The RyR2 staining and Cav1.2 staining are the same cell. Magnified images of the yellow boxes are shown in the middle row. Averaged fluorescent signals of all nanobars from multiple cells were averaged and displayed in the bottom row. n = 23, 23, 23, 11 cells for BFP-CAAX, JPH2, RyR2, and Cav1.2. Scale bar 10 µm in the top row, 2.5 µm in the middle row. l , Quantification of the ratio of the fluorescent intensity at the ends of nanobars (area in the magenta mask) over the average at the sides of nanobars (area in the green mask). Cell number is the same as in ( k ). ****P <0.0001; *P=0.0277. All experiments were independently replicated at least three times. All error bars represent STD. Kruskal-Wallis test corrected with Dunn’s multiple-comparison test was used to assess significance for ( g , l ).

    Journal: bioRxiv

    Article Title: Membrane Curvature Promotes ER-PM Contact Formation via Junctophilin-EHD Interactions

    doi: 10.1101/2024.06.29.601287

    Figure Lengend Snippet: a , SEM images of vertical nanopillars. Scale bar 1 µm on the left, 500 nm on the right. b , Schematics of the T-tubule system in a cardiomyocyte (left), ER-PM contacts formed at T-tubules (middle), and nanopillar-induced membrane invaginations (right). c-d , Co-Immunostained JPH2 (green) and α-actinin (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on flat areas (c) and on nanopillar areas (d). JPH2 (green) preferentially accumulates on nanopillar locations. Magnified images of the yellow boxes are shown at the bottom. Scale bar 10 µm on the top, 2.5 µm at the bottom. e , Averaged fluorescent signals for GFP-Sec61β, JPH2, and α-actinin. 101×101-pixel square regions of interest centered by each nanopillar were averaged and displayed. Over ∼3000 nanopillars were included in each condition. GFP-Sec61β n = 18 cells; JPH2 and α-actinin n = 21 cells. f , Intensity plots along the horizontal yellow line in e, normalized by the intensity value of the region between two nanopillars. Error bars represent standard deviation (STD) among values calculated from individual cells. g , The ratio of the fluorescent intensity at the nanopillars (pillar: area in the magenta mask) over the average intensity between the nanopillars (non-pillar: area inside the yellow mask). n=21, 10, 11, 18, 21 cells for JPH2, RyR2, Cav1.2, Sec61β, and α-actinin. ****P <0.0001; **P=0.0067; not-significant (NS) P>0.9999. h , Representative images of co-immunostained RyR2 (green) and Cav1.2 (red) in iPSC-CMs expressing GFP-Sec61β (magenta) on nanopillars. The solid arrowhead points at a cell with clear Cav1.2 signal on nanopillars and the hollow arrowhead points to a cell with minimal Cav1.2 on nanopillars. Scale bar 10 µm on the top, 2.5 µm on the bottom. i , Averaged fluorescent signals of ∼1200 nanopillars were included in each condition. n = 10 cells for RyR2; n = 11 cells for Cav1.2. j , SEM images of nanobars. Scale bar 5 µm on the top, 2 µm at the bottom. k , Representative images of iPSC-CMs with expressed BFP-CAAX, immunostaining of JPH2, RyR2, and Cav1.2 on nanobars. The RyR2 staining and Cav1.2 staining are the same cell. Magnified images of the yellow boxes are shown in the middle row. Averaged fluorescent signals of all nanobars from multiple cells were averaged and displayed in the bottom row. n = 23, 23, 23, 11 cells for BFP-CAAX, JPH2, RyR2, and Cav1.2. Scale bar 10 µm in the top row, 2.5 µm in the middle row. l , Quantification of the ratio of the fluorescent intensity at the ends of nanobars (area in the magenta mask) over the average at the sides of nanobars (area in the green mask). Cell number is the same as in ( k ). ****P <0.0001; *P=0.0277. All experiments were independently replicated at least three times. All error bars represent STD. Kruskal-Wallis test corrected with Dunn’s multiple-comparison test was used to assess significance for ( g , l ).

    Article Snippet: Anti-Junctophilin2 antibody (HPA052646), anti-α-actinin antibody (A7811), anti-Cav1.2 antibody (C1103), and anti-BIN1 antibody (SAB1408547) were purchased from Sigma.

    Techniques: Membrane, Expressing, Standard Deviation, Immunostaining, Staining, Comparison

    a , An illustration of the overlapping between the PM targeting and the curvature sensing candidates in JPH2 interactome. b , Representative images of GFP-JPH3 on nanobars shRNA knockdown of scramble, Clathrin, CAVIN1, CAV1/2, or EHD1/2/4. ShRNAs sequences were fused with fluorescence proteins to identify knockdown cells. Bright field and the fluorescent protein expression are shown on the left. Zooms are the enlarged view of the region in yellow boxes. Averaged nanobar images from multiple cells are shown on the right. Cell number for average: scramble = 28, shCLTC (Clathrin heavy chain) = 23, shCAV1/2 = 53, shCAVIN1 = 30, shEHD1/2/4 = 55. Scale bar 10 µm in whole cell images, 5 µm in zoom-in images. c , Quantifications of GFP-JPH3 end-to-side ratios upon shRNA knockdowns. Conditions and cell numbers are the same as in b. ****P<0.0001, NS(shClathrin) P>0.9999, NS(shCAV1shCAV2) P>0.9999, NS(shCAVIN1) P=0.0592. d , The distribution of GFP-JPH3 on nanobars before and after 10 mM MβCD treatment at 37°C for 30 min. Scale bar 5 µm. Right: averaged images of all bars in a single cell. e , Quantifications of the nanobar end-to-side ratios for GFP-JPH3 in U2OS cells before and after the MβCD treatment. n = 15 cells for each group. f , Representative images of U2OS cells co-expressing EHD4-GFP/mCherry with indicated JPH3 mutants. Enlarged views are zoomed from the yellow boxes in whole cell image on the left. Scale bar 10 µm in whole cell images, 5 µm in zoom-in images. g , Pearson’s correlation coefficients (PCC) between coexpressed EHD4-GFP/mCherry and JPH3 mutants shown in ( f ). Cell number: mCherry-8MORN_LCR = 21, GFP-8MORN_LCR-αHelix = 17, mCherry-LCR = 19, GFP-8MORN-αHelix = 19. ****P < 0.0001, **P = 0.0024 (8MORN_LCR-αHlx vs. 8MORN-αHlx), **P = 0.0075 (8MORN_LCR vs. 8MORN-αHlx), *P = 0.0174 (8MORN-αHlx vs. LCR), NS P= 0.9995 (8MORN_LCR-αHlx vs. 8MORN_LCR). h , PCC for colocalization between mCherry-8MORN_LCR and EHD4-GFP or between mCherry-8MORN_LCR and anti-clathrin. n = 21 cells for each group. ****P<0.0001. i , Co-immunoprecipitation (IP) from HEK cells expressing EHD4-GFP and mCherry-8MORN_LCR or mCherry-LCR. GFP was pulled via GFP-antibody beads, and the immunoprecipitates were blotted with mCherry antibody. mCherry-8MORN_LCR, but not mCherry-LCR, was pulled together with EHD4-GFP. All experiments were independently replicated at least three times. All error bars represent STD. Kruskal–Wallis test corrected with Dunn’s multiple-comparison test was used for ( c ). Unpaired two-tailed Welch’s t test was used to assess significance for ( e , h ). Brown-Forsythe and Welch ANOVA tests was used to assess significance for ( g ).

    Journal: bioRxiv

    Article Title: Membrane Curvature Promotes ER-PM Contact Formation via Junctophilin-EHD Interactions

    doi: 10.1101/2024.06.29.601287

    Figure Lengend Snippet: a , An illustration of the overlapping between the PM targeting and the curvature sensing candidates in JPH2 interactome. b , Representative images of GFP-JPH3 on nanobars shRNA knockdown of scramble, Clathrin, CAVIN1, CAV1/2, or EHD1/2/4. ShRNAs sequences were fused with fluorescence proteins to identify knockdown cells. Bright field and the fluorescent protein expression are shown on the left. Zooms are the enlarged view of the region in yellow boxes. Averaged nanobar images from multiple cells are shown on the right. Cell number for average: scramble = 28, shCLTC (Clathrin heavy chain) = 23, shCAV1/2 = 53, shCAVIN1 = 30, shEHD1/2/4 = 55. Scale bar 10 µm in whole cell images, 5 µm in zoom-in images. c , Quantifications of GFP-JPH3 end-to-side ratios upon shRNA knockdowns. Conditions and cell numbers are the same as in b. ****P<0.0001, NS(shClathrin) P>0.9999, NS(shCAV1shCAV2) P>0.9999, NS(shCAVIN1) P=0.0592. d , The distribution of GFP-JPH3 on nanobars before and after 10 mM MβCD treatment at 37°C for 30 min. Scale bar 5 µm. Right: averaged images of all bars in a single cell. e , Quantifications of the nanobar end-to-side ratios for GFP-JPH3 in U2OS cells before and after the MβCD treatment. n = 15 cells for each group. f , Representative images of U2OS cells co-expressing EHD4-GFP/mCherry with indicated JPH3 mutants. Enlarged views are zoomed from the yellow boxes in whole cell image on the left. Scale bar 10 µm in whole cell images, 5 µm in zoom-in images. g , Pearson’s correlation coefficients (PCC) between coexpressed EHD4-GFP/mCherry and JPH3 mutants shown in ( f ). Cell number: mCherry-8MORN_LCR = 21, GFP-8MORN_LCR-αHelix = 17, mCherry-LCR = 19, GFP-8MORN-αHelix = 19. ****P < 0.0001, **P = 0.0024 (8MORN_LCR-αHlx vs. 8MORN-αHlx), **P = 0.0075 (8MORN_LCR vs. 8MORN-αHlx), *P = 0.0174 (8MORN-αHlx vs. LCR), NS P= 0.9995 (8MORN_LCR-αHlx vs. 8MORN_LCR). h , PCC for colocalization between mCherry-8MORN_LCR and EHD4-GFP or between mCherry-8MORN_LCR and anti-clathrin. n = 21 cells for each group. ****P<0.0001. i , Co-immunoprecipitation (IP) from HEK cells expressing EHD4-GFP and mCherry-8MORN_LCR or mCherry-LCR. GFP was pulled via GFP-antibody beads, and the immunoprecipitates were blotted with mCherry antibody. mCherry-8MORN_LCR, but not mCherry-LCR, was pulled together with EHD4-GFP. All experiments were independently replicated at least three times. All error bars represent STD. Kruskal–Wallis test corrected with Dunn’s multiple-comparison test was used for ( c ). Unpaired two-tailed Welch’s t test was used to assess significance for ( e , h ). Brown-Forsythe and Welch ANOVA tests was used to assess significance for ( g ).

    Article Snippet: Anti-Junctophilin2 antibody (HPA052646), anti-α-actinin antibody (A7811), anti-Cav1.2 antibody (C1103), and anti-BIN1 antibody (SAB1408547) were purchased from Sigma.

    Techniques: shRNA, Knockdown, Fluorescence, Expressing, Immunoprecipitation, Comparison, Two Tailed Test

    a , EHD4, CAVIN1, Caveolin1, and Caveolin2 all show preferential accumulations at the nanobar ends. Quantifications of the end-to-side ratios are shown on the right. Cell number: n =14 cells for Caveolin2, n = 15 cells for all other groups. ***P=0.0001, **P=0.0015; ****P<0.0001. b , mRNA transcript levels in U2OS cells (shown as normalized transcripts per million, nTPM), obtained from protein atlas database. Genes highlighted in gray: not identified in the JPH2 interactome but are closely related either genetically or functionally. Genes including EHD3, CAVIN2, CAVIN3, and CAVIN4 were not selected as knockdown targets in KD experiments because of their low TPM level in U2OS cells. c , Western blots confirming the effective shRNA knockdown of CAV1, CAV2, EHD1, EHD2, CLTC (clathrin heavy chain), and BIN1. GAPDH was blotted as a loading control. d , Representative images of U2OS cells co-expressing EHD4-mCherry with EHD1-GFP (Top) or with EHD2-GFP (Bottom). Enlarged views are zoomed from the yellow boxes in the whole cell images. e , Representative images of U2OS cells expressing GFP-JPH3 (green) and EHD4-mCh (magenta) seeding on nanobars. Enlarged views are zoomed from the yellow boxes in whole cell images. f , Representative images of immunostaining of EHD2 or EHD4 in iPSC-CMs or rat embryonic CMs on nanopillars. Magnified images of the region in yellow boxes are shown in the bottom row. Scale bar 10 µm in the top row, 2.5 µm in the bottom row. g , Representative images of immunostaining of EHD4 in iPSC-CMs on nanobars. Magnified images of the region in yellow boxes are shown in the middle row. Averaged fluorescent signals of all nanobars from multiple cells were averaged and displayed in the bottom row. n = 16 cells for the average image. Bottom graph: quantification of the end to side ratios of EHD4 compared to that of BFP-CAAX expressed in iPSC-CM on nanobars. Cell number: CAAX = 23 (same data as in ), EHD4 = 16. ****P <0.0001. h , Representative images of immunostaining of JPH2 in iPSC cells transfected with shRNA of scramble (top) or EHD1/2/4 (bottom). From left to right: Bright field image, fluorescent protein expression indicating successful transfection of shRNA, and JPH2 staining. Averaged nanobar images of JPH2 staining from multiple cells are shown at the bottom of JPH staining panels. Quantifications of the end-to-side ratios are shown on the right. Cell number for both average and quantification: Scramble = 24, shEHD1/2/4 = 26. ****P <0.0001. Scale bars 10 µm in all whole cell images, 5 µm in all zoomed images unless otherwise mentioned. All experiments were independently replicated at least two times. All error bars represent STD. Kruskal–Wallis test corrected with Dunn’s multiple-comparison test was used to assess significance in ( a ). Unpaired two-tailed Welch’s t test was used to assess significance in ( g ). Two-tailed Mann-Whitney test was used to assess significance in ( h ).

    Journal: bioRxiv

    Article Title: Membrane Curvature Promotes ER-PM Contact Formation via Junctophilin-EHD Interactions

    doi: 10.1101/2024.06.29.601287

    Figure Lengend Snippet: a , EHD4, CAVIN1, Caveolin1, and Caveolin2 all show preferential accumulations at the nanobar ends. Quantifications of the end-to-side ratios are shown on the right. Cell number: n =14 cells for Caveolin2, n = 15 cells for all other groups. ***P=0.0001, **P=0.0015; ****P<0.0001. b , mRNA transcript levels in U2OS cells (shown as normalized transcripts per million, nTPM), obtained from protein atlas database. Genes highlighted in gray: not identified in the JPH2 interactome but are closely related either genetically or functionally. Genes including EHD3, CAVIN2, CAVIN3, and CAVIN4 were not selected as knockdown targets in KD experiments because of their low TPM level in U2OS cells. c , Western blots confirming the effective shRNA knockdown of CAV1, CAV2, EHD1, EHD2, CLTC (clathrin heavy chain), and BIN1. GAPDH was blotted as a loading control. d , Representative images of U2OS cells co-expressing EHD4-mCherry with EHD1-GFP (Top) or with EHD2-GFP (Bottom). Enlarged views are zoomed from the yellow boxes in the whole cell images. e , Representative images of U2OS cells expressing GFP-JPH3 (green) and EHD4-mCh (magenta) seeding on nanobars. Enlarged views are zoomed from the yellow boxes in whole cell images. f , Representative images of immunostaining of EHD2 or EHD4 in iPSC-CMs or rat embryonic CMs on nanopillars. Magnified images of the region in yellow boxes are shown in the bottom row. Scale bar 10 µm in the top row, 2.5 µm in the bottom row. g , Representative images of immunostaining of EHD4 in iPSC-CMs on nanobars. Magnified images of the region in yellow boxes are shown in the middle row. Averaged fluorescent signals of all nanobars from multiple cells were averaged and displayed in the bottom row. n = 16 cells for the average image. Bottom graph: quantification of the end to side ratios of EHD4 compared to that of BFP-CAAX expressed in iPSC-CM on nanobars. Cell number: CAAX = 23 (same data as in ), EHD4 = 16. ****P <0.0001. h , Representative images of immunostaining of JPH2 in iPSC cells transfected with shRNA of scramble (top) or EHD1/2/4 (bottom). From left to right: Bright field image, fluorescent protein expression indicating successful transfection of shRNA, and JPH2 staining. Averaged nanobar images of JPH2 staining from multiple cells are shown at the bottom of JPH staining panels. Quantifications of the end-to-side ratios are shown on the right. Cell number for both average and quantification: Scramble = 24, shEHD1/2/4 = 26. ****P <0.0001. Scale bars 10 µm in all whole cell images, 5 µm in all zoomed images unless otherwise mentioned. All experiments were independently replicated at least two times. All error bars represent STD. Kruskal–Wallis test corrected with Dunn’s multiple-comparison test was used to assess significance in ( a ). Unpaired two-tailed Welch’s t test was used to assess significance in ( g ). Two-tailed Mann-Whitney test was used to assess significance in ( h ).

    Article Snippet: Anti-Junctophilin2 antibody (HPA052646), anti-α-actinin antibody (A7811), anti-Cav1.2 antibody (C1103), and anti-BIN1 antibody (SAB1408547) were purchased from Sigma.

    Techniques: Knockdown, Western Blot, shRNA, Control, Expressing, Immunostaining, Transfection, Staining, Comparison, Two Tailed Test, MANN-WHITNEY