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polyclonal rabbit anti-dsg2 for sted  (Progen Biotechnik)

 
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    Progen Biotechnik polyclonal rabbit anti-dsg2 for sted
    (A) Modeling of the potential cis interaction between 2 <t>Dsg2</t> extracellular 1 domains (EC1s). Dashed line indicates 2-fold symmetry axis of the 2 EC1s. (B) Structure of the monomeric peptide (Dsg2-IP) fitted to the EC1 of Dsg2. Residues potentially in contact with the peptide are shown as sticks and numbered in 3-letter code accordingly. (C) Magnification of B. Dsg2-IP shown; C atoms colored in cyan. Interface residues located on the central β-sheet are shown as sticks with C atoms colored in green. (D and F) Representative time course of duration of R-R intervals of perfused hearts from Pg-KO mice with AC-like phenotype under baseline conditions and after addition of Dsg2-LP. Every dot indicates the duration of 1 beat-to-beat interval plotted against perfusion time. VE-Cad-LP served as peptide control. Right panel shows representative ECG traces from indicated time points. (E and G) Analysis of SDNN derived from Pg-KO hearts treated as described in D and F. (H and I) Representative time course of duration of R-R intervals with analysis of the SDNN of perfused hearts from WT mice under baseline conditions and after addition of Dsg2-LP (n = 5 mice for Dsg-LP and Pg-KO and WT; n = 3 mice for VE-Cad-LP) n.s., P ≥ 0.05, *P < 0.05. Two-tailed paired Student’s t test with 95% confidence level was performed. Black lines indicate paired values.
    Polyclonal Rabbit Anti Dsg2 For Sted, supplied by Progen Biotechnik, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polyclonal rabbit anti-dsg2 for sted/product/Progen Biotechnik
    Average 90 stars, based on 1 article reviews
    polyclonal rabbit anti-dsg2 for sted - by Bioz Stars, 2026-02
    90/100 stars

    Images

    1) Product Images from "Stabilization of desmoglein-2 binding rescues arrhythmia in arrhythmogenic cardiomyopathy"

    Article Title: Stabilization of desmoglein-2 binding rescues arrhythmia in arrhythmogenic cardiomyopathy

    Journal: JCI Insight

    doi: 10.1172/jci.insight.130141

    (A) Modeling of the potential cis interaction between 2 Dsg2 extracellular 1 domains (EC1s). Dashed line indicates 2-fold symmetry axis of the 2 EC1s. (B) Structure of the monomeric peptide (Dsg2-IP) fitted to the EC1 of Dsg2. Residues potentially in contact with the peptide are shown as sticks and numbered in 3-letter code accordingly. (C) Magnification of B. Dsg2-IP shown; C atoms colored in cyan. Interface residues located on the central β-sheet are shown as sticks with C atoms colored in green. (D and F) Representative time course of duration of R-R intervals of perfused hearts from Pg-KO mice with AC-like phenotype under baseline conditions and after addition of Dsg2-LP. Every dot indicates the duration of 1 beat-to-beat interval plotted against perfusion time. VE-Cad-LP served as peptide control. Right panel shows representative ECG traces from indicated time points. (E and G) Analysis of SDNN derived from Pg-KO hearts treated as described in D and F. (H and I) Representative time course of duration of R-R intervals with analysis of the SDNN of perfused hearts from WT mice under baseline conditions and after addition of Dsg2-LP (n = 5 mice for Dsg-LP and Pg-KO and WT; n = 3 mice for VE-Cad-LP) n.s., P ≥ 0.05, *P < 0.05. Two-tailed paired Student’s t test with 95% confidence level was performed. Black lines indicate paired values.
    Figure Legend Snippet: (A) Modeling of the potential cis interaction between 2 Dsg2 extracellular 1 domains (EC1s). Dashed line indicates 2-fold symmetry axis of the 2 EC1s. (B) Structure of the monomeric peptide (Dsg2-IP) fitted to the EC1 of Dsg2. Residues potentially in contact with the peptide are shown as sticks and numbered in 3-letter code accordingly. (C) Magnification of B. Dsg2-IP shown; C atoms colored in cyan. Interface residues located on the central β-sheet are shown as sticks with C atoms colored in green. (D and F) Representative time course of duration of R-R intervals of perfused hearts from Pg-KO mice with AC-like phenotype under baseline conditions and after addition of Dsg2-LP. Every dot indicates the duration of 1 beat-to-beat interval plotted against perfusion time. VE-Cad-LP served as peptide control. Right panel shows representative ECG traces from indicated time points. (E and G) Analysis of SDNN derived from Pg-KO hearts treated as described in D and F. (H and I) Representative time course of duration of R-R intervals with analysis of the SDNN of perfused hearts from WT mice under baseline conditions and after addition of Dsg2-LP (n = 5 mice for Dsg-LP and Pg-KO and WT; n = 3 mice for VE-Cad-LP) n.s., P ≥ 0.05, *P < 0.05. Two-tailed paired Student’s t test with 95% confidence level was performed. Black lines indicate paired values.

    Techniques Used: Derivative Assay, Two Tailed Test

    (A) STED images of ICDs of WT and Pg-KO mice treated with Dsg2-LP for 10 minutes and stained for Dsg2 (red) and Cx43 (green) (4 ICDs per mouse were analyzed for quantification; n = 3 mice per condition). Scale bar: 2 μm. Colocalization of both proteins was analyzed, and colocalized pixels are visualized in white. (B) Quantification of colocalized pixels. *P < 0.05. Repeated-measures 2-way ANOVA with Holm-Šidák post hoc test was performed.
    Figure Legend Snippet: (A) STED images of ICDs of WT and Pg-KO mice treated with Dsg2-LP for 10 minutes and stained for Dsg2 (red) and Cx43 (green) (4 ICDs per mouse were analyzed for quantification; n = 3 mice per condition). Scale bar: 2 μm. Colocalization of both proteins was analyzed, and colocalized pixels are visualized in white. (B) Quantification of colocalized pixels. *P < 0.05. Repeated-measures 2-way ANOVA with Holm-Šidák post hoc test was performed.

    Techniques Used: Staining

    (A) Dissociation assays of HL-1 monolayer treated with Dsg2- or Pg-siRNA and additional Dsg2-LP treatment (n = 8). One-way ANOVA with Bonferroni’s post hoc test. (B) Western blot analysis for Dsg2 and Pg performed in parallel to dissociation assays to show successful protein reduction after Dsg2- or Pg-siRNA treatment. α-Tubulin (α-Tub) served as loading control (n = 8). (C) Dissociation assay of HL-1 cell monolayer after disruption of cell cohesion by Trp or Dsg2-IP with Dsg2-LP treatment (control n = 8; Trp n = 8; Trp + Dsg-LP n = 7; Dsg2-LP n = 5; Dsg2-IP n = 3). One-way ANOVA with Bonferroni’s post hoc test. (D) Representative images of bead fragmentation after application of mechanical stress. Adhesion assay of Dsg2-Fc–coated beads with same conditions as C. (E) Ca2+ chelator EGTA and anti-Dsg2 antibody served as negative controls (control n = 10; EGTA n = 9; anti-Dsg2 n = 10; Trp n = 8; Trp + Dsg-LP n = 8; Dsg2-LP n = 8; Dsg2-IP n = 8). One-way ANOVA with Bonferroni’s post hoc test. (F) Representative immunostaining of HL-1 cells stained for Dsg2 (red) and Cx43 (green) treated with Trp, Dsg2-LP, or a combination of both for 24 hours (n = 8); scale bar: 10 μm. (G) Quantitative analysis of single-Dsg2 particle size (>30 cells from 3–4 independent experiments). (H) Quantitative analysis of Cx43 staining as ratio of intensity of staining at cell-cell border versus cytosol (>30 cells from 3 independent experiments). *P < 0.05. One-way ANOVA with Tukey’s post hoc test.
    Figure Legend Snippet: (A) Dissociation assays of HL-1 monolayer treated with Dsg2- or Pg-siRNA and additional Dsg2-LP treatment (n = 8). One-way ANOVA with Bonferroni’s post hoc test. (B) Western blot analysis for Dsg2 and Pg performed in parallel to dissociation assays to show successful protein reduction after Dsg2- or Pg-siRNA treatment. α-Tubulin (α-Tub) served as loading control (n = 8). (C) Dissociation assay of HL-1 cell monolayer after disruption of cell cohesion by Trp or Dsg2-IP with Dsg2-LP treatment (control n = 8; Trp n = 8; Trp + Dsg-LP n = 7; Dsg2-LP n = 5; Dsg2-IP n = 3). One-way ANOVA with Bonferroni’s post hoc test. (D) Representative images of bead fragmentation after application of mechanical stress. Adhesion assay of Dsg2-Fc–coated beads with same conditions as C. (E) Ca2+ chelator EGTA and anti-Dsg2 antibody served as negative controls (control n = 10; EGTA n = 9; anti-Dsg2 n = 10; Trp n = 8; Trp + Dsg-LP n = 8; Dsg2-LP n = 8; Dsg2-IP n = 8). One-way ANOVA with Bonferroni’s post hoc test. (F) Representative immunostaining of HL-1 cells stained for Dsg2 (red) and Cx43 (green) treated with Trp, Dsg2-LP, or a combination of both for 24 hours (n = 8); scale bar: 10 μm. (G) Quantitative analysis of single-Dsg2 particle size (>30 cells from 3–4 independent experiments). (H) Quantitative analysis of Cx43 staining as ratio of intensity of staining at cell-cell border versus cytosol (>30 cells from 3 independent experiments). *P < 0.05. One-way ANOVA with Tukey’s post hoc test.

    Techniques Used: Western Blot, Cell Adhesion Assay, Immunostaining, Staining

    (A) Representative MEA heatmaps of transversal slices of WT and Pg-KO hearts treated with Dsg2-LP for 1 hour showing the delay of spontaneous excitations relative to a reference electrode (*). Colors correspond to the delay times of excitation given in the scale. Distance between neighboring electrodes: 200 μm. (B) Corresponding analysis of conduction velocity measured in cardiac slices as shown in A (WT control n = 9; WT Dsg2-LP n = 6 from 3 mice; Pg-KO control n = 11; Pg-KO Dsg2-LP n = 12 from 5 mice). Two-way ANOVA with Holm-Šidák post hoc test was performed. (C and F) MEA heatmaps from spontaneously contracting HL-1 cardiomyocytes treated with Dsg2-LP. Pg and Dsg2-levels were reduced by siRNA (Pg-siRNA, Dsg2-siRNA; n.t.-siRNA served as control). Distance between neighboring electrodes: 200 μm. (D, E, G, and H) Corresponding analysis of conduction velocity and SDNN of MEAs as described in C (control n = 5; Trp n = 5; Trp + Dsg2-LP n = 4; Dsg2-LP n = 5), and (F) (n.t.-siRNA n = 5; Pg-siRNA n = 4; Pg-siRNA + Dsg2-LP n = 3; Dsg2-siRNA n = 5; Dsg2-siRNA + Dsg2-LP n = 3). One-way ANOVA with Bonferroni’s post hoc test was performed. (I and J) Western blot analysis shows phosphorylation of Cx43 at serine 368 (pCx43 Ser368) in response to PKC activation by PMA and PKC inhibition by Bim-X or Dsg2-LP in HL-1 (I) and WT mice ventricular cardiac slices (J). Values in top row display mean of densitometric analysis ± SD. Total Cx43 was used as a reference protein. α-Tubulin (α-Tub) served as loading control (n = 4). (K–M) MEAs of Pg-reduced HL-1 cardiomyocytes treated with Bim-X and Dsg2-LP with corresponding analysis of conduction velocity and SDNN (n = 3). *P < 0.05. One-way ANOVA with Bonferroni’s post hoc test.
    Figure Legend Snippet: (A) Representative MEA heatmaps of transversal slices of WT and Pg-KO hearts treated with Dsg2-LP for 1 hour showing the delay of spontaneous excitations relative to a reference electrode (*). Colors correspond to the delay times of excitation given in the scale. Distance between neighboring electrodes: 200 μm. (B) Corresponding analysis of conduction velocity measured in cardiac slices as shown in A (WT control n = 9; WT Dsg2-LP n = 6 from 3 mice; Pg-KO control n = 11; Pg-KO Dsg2-LP n = 12 from 5 mice). Two-way ANOVA with Holm-Šidák post hoc test was performed. (C and F) MEA heatmaps from spontaneously contracting HL-1 cardiomyocytes treated with Dsg2-LP. Pg and Dsg2-levels were reduced by siRNA (Pg-siRNA, Dsg2-siRNA; n.t.-siRNA served as control). Distance between neighboring electrodes: 200 μm. (D, E, G, and H) Corresponding analysis of conduction velocity and SDNN of MEAs as described in C (control n = 5; Trp n = 5; Trp + Dsg2-LP n = 4; Dsg2-LP n = 5), and (F) (n.t.-siRNA n = 5; Pg-siRNA n = 4; Pg-siRNA + Dsg2-LP n = 3; Dsg2-siRNA n = 5; Dsg2-siRNA + Dsg2-LP n = 3). One-way ANOVA with Bonferroni’s post hoc test was performed. (I and J) Western blot analysis shows phosphorylation of Cx43 at serine 368 (pCx43 Ser368) in response to PKC activation by PMA and PKC inhibition by Bim-X or Dsg2-LP in HL-1 (I) and WT mice ventricular cardiac slices (J). Values in top row display mean of densitometric analysis ± SD. Total Cx43 was used as a reference protein. α-Tubulin (α-Tub) served as loading control (n = 4). (K–M) MEAs of Pg-reduced HL-1 cardiomyocytes treated with Bim-X and Dsg2-LP with corresponding analysis of conduction velocity and SDNN (n = 3). *P < 0.05. One-way ANOVA with Bonferroni’s post hoc test.

    Techniques Used: Western Blot, Activation Assay, Inhibition

    (A) Schematic of atomic force microscope (AFM) force mapping setup. The tip of a cantilever functionalized with the extracellular domains of adhesion molecules via a flexible PEG linker (step 1) is repeatedly extended to (step 2) and retracted from (step 3) a functionalized surface. Interaction of the tip- and surface-bound proteins is detectable by cantilever deflection. (B and C) Analysis of unbinding force and binding frequency of cell-free AFM measurements with Dsg2- or N-Cad–coated tips probed on mica sheets with respective coatings. Every dot in the right panel represents the mean value of 1 independent experiment, bars indicate mean ± SD, and black lines connect paired values. n = 6 for Dsg2; n = 5 for N-Cad. n.s., P ≥ 0.05, *P < 0.05. Two-tailed paired Student’s t test.
    Figure Legend Snippet: (A) Schematic of atomic force microscope (AFM) force mapping setup. The tip of a cantilever functionalized with the extracellular domains of adhesion molecules via a flexible PEG linker (step 1) is repeatedly extended to (step 2) and retracted from (step 3) a functionalized surface. Interaction of the tip- and surface-bound proteins is detectable by cantilever deflection. (B and C) Analysis of unbinding force and binding frequency of cell-free AFM measurements with Dsg2- or N-Cad–coated tips probed on mica sheets with respective coatings. Every dot in the right panel represents the mean value of 1 independent experiment, bars indicate mean ± SD, and black lines connect paired values. n = 6 for Dsg2; n = 5 for N-Cad. n.s., P ≥ 0.05, *P < 0.05. Two-tailed paired Student’s t test.

    Techniques Used: Microscopy, Binding Assay, Two Tailed Test

    Representative binding event and topography images of AFM force mapping performed with a Dsg2-coated tip on HL-1 cardiomyocytes (A) or cardiomyocytes isolated from Dsg2-KO mice or WT littermates (D, G, and J) treated with Dsg2-LP 30–90 minutes. Cyan line indicates cell-cell border. Every white pixel represents 1 binding event at the respective location. (A) n = 5 independent experiments, scale bar: 1 μm. (D) n = 7 mice, (G) n = 6 mice per phenotype, and (J) n = 6 mice; scale bar: 2 μm. Unbinding forces (B, E, H, and K) and binding frequency (C, F, I, and L) of Dsg2-mediated binding events corresponding to respective panels on the left. Every dot represents the mean value of 1 independent experiment, bars indicate mean ± SD, and black lines connect paired values. *P < 0.05. Two-tailed paired Student’s t test was performed.
    Figure Legend Snippet: Representative binding event and topography images of AFM force mapping performed with a Dsg2-coated tip on HL-1 cardiomyocytes (A) or cardiomyocytes isolated from Dsg2-KO mice or WT littermates (D, G, and J) treated with Dsg2-LP 30–90 minutes. Cyan line indicates cell-cell border. Every white pixel represents 1 binding event at the respective location. (A) n = 5 independent experiments, scale bar: 1 μm. (D) n = 7 mice, (G) n = 6 mice per phenotype, and (J) n = 6 mice; scale bar: 2 μm. Unbinding forces (B, E, H, and K) and binding frequency (C, F, I, and L) of Dsg2-mediated binding events corresponding to respective panels on the left. Every dot represents the mean value of 1 independent experiment, bars indicate mean ± SD, and black lines connect paired values. *P < 0.05. Two-tailed paired Student’s t test was performed.

    Techniques Used: Binding Assay, Isolation, Two Tailed Test

    (A) Membrane-impermeable cross-linking with sulfo-EGS performed after 24-hour treatment with Dsg2-LP in HL-1 cells. Western blot analyses were done for Dsg2 and N-Cad. α-Tubulin (α-Tub) served as loading control. Representative image of 4 repeated experiments. (B) Densitometric quantification of Dsg2 and N-Cad bands showing the ratio of cross-linked proteins to total protein. n = 4. *P < 0.05. Two-tailed paired Student’s t test with 95% confidence levels was performed.
    Figure Legend Snippet: (A) Membrane-impermeable cross-linking with sulfo-EGS performed after 24-hour treatment with Dsg2-LP in HL-1 cells. Western blot analyses were done for Dsg2 and N-Cad. α-Tubulin (α-Tub) served as loading control. Representative image of 4 repeated experiments. (B) Densitometric quantification of Dsg2 and N-Cad bands showing the ratio of cross-linked proteins to total protein. n = 4. *P < 0.05. Two-tailed paired Student’s t test with 95% confidence levels was performed.

    Techniques Used: Western Blot, Two Tailed Test



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    polyclonal rabbit anti-dsg2 for sted  (Progen Biotechnik)
    90
    Progen Biotechnik polyclonal rabbit anti-dsg2 for sted
    (A) Modeling of the potential cis interaction between 2 <t>Dsg2</t> extracellular 1 domains (EC1s). Dashed line indicates 2-fold symmetry axis of the 2 EC1s. (B) Structure of the monomeric peptide (Dsg2-IP) fitted to the EC1 of Dsg2. Residues potentially in contact with the peptide are shown as sticks and numbered in 3-letter code accordingly. (C) Magnification of B. Dsg2-IP shown; C atoms colored in cyan. Interface residues located on the central β-sheet are shown as sticks with C atoms colored in green. (D and F) Representative time course of duration of R-R intervals of perfused hearts from Pg-KO mice with AC-like phenotype under baseline conditions and after addition of Dsg2-LP. Every dot indicates the duration of 1 beat-to-beat interval plotted against perfusion time. VE-Cad-LP served as peptide control. Right panel shows representative ECG traces from indicated time points. (E and G) Analysis of SDNN derived from Pg-KO hearts treated as described in D and F. (H and I) Representative time course of duration of R-R intervals with analysis of the SDNN of perfused hearts from WT mice under baseline conditions and after addition of Dsg2-LP (n = 5 mice for Dsg-LP and Pg-KO and WT; n = 3 mice for VE-Cad-LP) n.s., P ≥ 0.05, *P < 0.05. Two-tailed paired Student’s t test with 95% confidence level was performed. Black lines indicate paired values.
    Polyclonal Rabbit Anti Dsg2 For Sted, supplied by Progen Biotechnik, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polyclonal rabbit anti-dsg2 for sted/product/Progen Biotechnik
    Average 90 stars, based on 1 article reviews
    polyclonal rabbit anti-dsg2 for sted - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier

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    (A) Modeling of the potential cis interaction between 2 Dsg2 extracellular 1 domains (EC1s). Dashed line indicates 2-fold symmetry axis of the 2 EC1s. (B) Structure of the monomeric peptide (Dsg2-IP) fitted to the EC1 of Dsg2. Residues potentially in contact with the peptide are shown as sticks and numbered in 3-letter code accordingly. (C) Magnification of B. Dsg2-IP shown; C atoms colored in cyan. Interface residues located on the central β-sheet are shown as sticks with C atoms colored in green. (D and F) Representative time course of duration of R-R intervals of perfused hearts from Pg-KO mice with AC-like phenotype under baseline conditions and after addition of Dsg2-LP. Every dot indicates the duration of 1 beat-to-beat interval plotted against perfusion time. VE-Cad-LP served as peptide control. Right panel shows representative ECG traces from indicated time points. (E and G) Analysis of SDNN derived from Pg-KO hearts treated as described in D and F. (H and I) Representative time course of duration of R-R intervals with analysis of the SDNN of perfused hearts from WT mice under baseline conditions and after addition of Dsg2-LP (n = 5 mice for Dsg-LP and Pg-KO and WT; n = 3 mice for VE-Cad-LP) n.s., P ≥ 0.05, *P < 0.05. Two-tailed paired Student’s t test with 95% confidence level was performed. Black lines indicate paired values.

    Journal: JCI Insight

    Article Title: Stabilization of desmoglein-2 binding rescues arrhythmia in arrhythmogenic cardiomyopathy

    doi: 10.1172/jci.insight.130141

    Figure Lengend Snippet: (A) Modeling of the potential cis interaction between 2 Dsg2 extracellular 1 domains (EC1s). Dashed line indicates 2-fold symmetry axis of the 2 EC1s. (B) Structure of the monomeric peptide (Dsg2-IP) fitted to the EC1 of Dsg2. Residues potentially in contact with the peptide are shown as sticks and numbered in 3-letter code accordingly. (C) Magnification of B. Dsg2-IP shown; C atoms colored in cyan. Interface residues located on the central β-sheet are shown as sticks with C atoms colored in green. (D and F) Representative time course of duration of R-R intervals of perfused hearts from Pg-KO mice with AC-like phenotype under baseline conditions and after addition of Dsg2-LP. Every dot indicates the duration of 1 beat-to-beat interval plotted against perfusion time. VE-Cad-LP served as peptide control. Right panel shows representative ECG traces from indicated time points. (E and G) Analysis of SDNN derived from Pg-KO hearts treated as described in D and F. (H and I) Representative time course of duration of R-R intervals with analysis of the SDNN of perfused hearts from WT mice under baseline conditions and after addition of Dsg2-LP (n = 5 mice for Dsg-LP and Pg-KO and WT; n = 3 mice for VE-Cad-LP) n.s., P ≥ 0.05, *P < 0.05. Two-tailed paired Student’s t test with 95% confidence level was performed. Black lines indicate paired values.

    Article Snippet: The following primary antibodies were used for Western blot (WB) analysis or immunostaining evaluated by confocal or STED microscopy: monoclonal mouse anti-Dsg1/2 for WB and confocal microscopy (Progen Biotechnik, 61002), polyclonal rabbit anti-Dsg2 for STED (Progen Biotechnik, 610121), monoclonal mouse anti-Pg for WB and confocal microscopy (Progen Biotechnik, 61005), monoclonal mouse anti–α-tubulin for WB (Abcam, ab7291), polyclonal rabbit anti-Cx43 for confocal microscopy and WB (MilliporeSigma, SAB4501175), monoclonal mouse anti-Cx43 for STED (Merck KGaA, MAB3067), phospho-Cx43 Ser368 for WB (Santa Cruz Biotechnology Inc., sc-101660-r), monoclonal mouse anti–N-cadherin (BD Biosciences, 610921), and monoclonal mouse anti-Dp (Progen Biotechnik, 61003).

    Techniques: Derivative Assay, Two Tailed Test

    (A) STED images of ICDs of WT and Pg-KO mice treated with Dsg2-LP for 10 minutes and stained for Dsg2 (red) and Cx43 (green) (4 ICDs per mouse were analyzed for quantification; n = 3 mice per condition). Scale bar: 2 μm. Colocalization of both proteins was analyzed, and colocalized pixels are visualized in white. (B) Quantification of colocalized pixels. *P < 0.05. Repeated-measures 2-way ANOVA with Holm-Šidák post hoc test was performed.

    Journal: JCI Insight

    Article Title: Stabilization of desmoglein-2 binding rescues arrhythmia in arrhythmogenic cardiomyopathy

    doi: 10.1172/jci.insight.130141

    Figure Lengend Snippet: (A) STED images of ICDs of WT and Pg-KO mice treated with Dsg2-LP for 10 minutes and stained for Dsg2 (red) and Cx43 (green) (4 ICDs per mouse were analyzed for quantification; n = 3 mice per condition). Scale bar: 2 μm. Colocalization of both proteins was analyzed, and colocalized pixels are visualized in white. (B) Quantification of colocalized pixels. *P < 0.05. Repeated-measures 2-way ANOVA with Holm-Šidák post hoc test was performed.

    Article Snippet: The following primary antibodies were used for Western blot (WB) analysis or immunostaining evaluated by confocal or STED microscopy: monoclonal mouse anti-Dsg1/2 for WB and confocal microscopy (Progen Biotechnik, 61002), polyclonal rabbit anti-Dsg2 for STED (Progen Biotechnik, 610121), monoclonal mouse anti-Pg for WB and confocal microscopy (Progen Biotechnik, 61005), monoclonal mouse anti–α-tubulin for WB (Abcam, ab7291), polyclonal rabbit anti-Cx43 for confocal microscopy and WB (MilliporeSigma, SAB4501175), monoclonal mouse anti-Cx43 for STED (Merck KGaA, MAB3067), phospho-Cx43 Ser368 for WB (Santa Cruz Biotechnology Inc., sc-101660-r), monoclonal mouse anti–N-cadherin (BD Biosciences, 610921), and monoclonal mouse anti-Dp (Progen Biotechnik, 61003).

    Techniques: Staining

    (A) Dissociation assays of HL-1 monolayer treated with Dsg2- or Pg-siRNA and additional Dsg2-LP treatment (n = 8). One-way ANOVA with Bonferroni’s post hoc test. (B) Western blot analysis for Dsg2 and Pg performed in parallel to dissociation assays to show successful protein reduction after Dsg2- or Pg-siRNA treatment. α-Tubulin (α-Tub) served as loading control (n = 8). (C) Dissociation assay of HL-1 cell monolayer after disruption of cell cohesion by Trp or Dsg2-IP with Dsg2-LP treatment (control n = 8; Trp n = 8; Trp + Dsg-LP n = 7; Dsg2-LP n = 5; Dsg2-IP n = 3). One-way ANOVA with Bonferroni’s post hoc test. (D) Representative images of bead fragmentation after application of mechanical stress. Adhesion assay of Dsg2-Fc–coated beads with same conditions as C. (E) Ca2+ chelator EGTA and anti-Dsg2 antibody served as negative controls (control n = 10; EGTA n = 9; anti-Dsg2 n = 10; Trp n = 8; Trp + Dsg-LP n = 8; Dsg2-LP n = 8; Dsg2-IP n = 8). One-way ANOVA with Bonferroni’s post hoc test. (F) Representative immunostaining of HL-1 cells stained for Dsg2 (red) and Cx43 (green) treated with Trp, Dsg2-LP, or a combination of both for 24 hours (n = 8); scale bar: 10 μm. (G) Quantitative analysis of single-Dsg2 particle size (>30 cells from 3–4 independent experiments). (H) Quantitative analysis of Cx43 staining as ratio of intensity of staining at cell-cell border versus cytosol (>30 cells from 3 independent experiments). *P < 0.05. One-way ANOVA with Tukey’s post hoc test.

    Journal: JCI Insight

    Article Title: Stabilization of desmoglein-2 binding rescues arrhythmia in arrhythmogenic cardiomyopathy

    doi: 10.1172/jci.insight.130141

    Figure Lengend Snippet: (A) Dissociation assays of HL-1 monolayer treated with Dsg2- or Pg-siRNA and additional Dsg2-LP treatment (n = 8). One-way ANOVA with Bonferroni’s post hoc test. (B) Western blot analysis for Dsg2 and Pg performed in parallel to dissociation assays to show successful protein reduction after Dsg2- or Pg-siRNA treatment. α-Tubulin (α-Tub) served as loading control (n = 8). (C) Dissociation assay of HL-1 cell monolayer after disruption of cell cohesion by Trp or Dsg2-IP with Dsg2-LP treatment (control n = 8; Trp n = 8; Trp + Dsg-LP n = 7; Dsg2-LP n = 5; Dsg2-IP n = 3). One-way ANOVA with Bonferroni’s post hoc test. (D) Representative images of bead fragmentation after application of mechanical stress. Adhesion assay of Dsg2-Fc–coated beads with same conditions as C. (E) Ca2+ chelator EGTA and anti-Dsg2 antibody served as negative controls (control n = 10; EGTA n = 9; anti-Dsg2 n = 10; Trp n = 8; Trp + Dsg-LP n = 8; Dsg2-LP n = 8; Dsg2-IP n = 8). One-way ANOVA with Bonferroni’s post hoc test. (F) Representative immunostaining of HL-1 cells stained for Dsg2 (red) and Cx43 (green) treated with Trp, Dsg2-LP, or a combination of both for 24 hours (n = 8); scale bar: 10 μm. (G) Quantitative analysis of single-Dsg2 particle size (>30 cells from 3–4 independent experiments). (H) Quantitative analysis of Cx43 staining as ratio of intensity of staining at cell-cell border versus cytosol (>30 cells from 3 independent experiments). *P < 0.05. One-way ANOVA with Tukey’s post hoc test.

    Article Snippet: The following primary antibodies were used for Western blot (WB) analysis or immunostaining evaluated by confocal or STED microscopy: monoclonal mouse anti-Dsg1/2 for WB and confocal microscopy (Progen Biotechnik, 61002), polyclonal rabbit anti-Dsg2 for STED (Progen Biotechnik, 610121), monoclonal mouse anti-Pg for WB and confocal microscopy (Progen Biotechnik, 61005), monoclonal mouse anti–α-tubulin for WB (Abcam, ab7291), polyclonal rabbit anti-Cx43 for confocal microscopy and WB (MilliporeSigma, SAB4501175), monoclonal mouse anti-Cx43 for STED (Merck KGaA, MAB3067), phospho-Cx43 Ser368 for WB (Santa Cruz Biotechnology Inc., sc-101660-r), monoclonal mouse anti–N-cadherin (BD Biosciences, 610921), and monoclonal mouse anti-Dp (Progen Biotechnik, 61003).

    Techniques: Western Blot, Cell Adhesion Assay, Immunostaining, Staining

    (A) Representative MEA heatmaps of transversal slices of WT and Pg-KO hearts treated with Dsg2-LP for 1 hour showing the delay of spontaneous excitations relative to a reference electrode (*). Colors correspond to the delay times of excitation given in the scale. Distance between neighboring electrodes: 200 μm. (B) Corresponding analysis of conduction velocity measured in cardiac slices as shown in A (WT control n = 9; WT Dsg2-LP n = 6 from 3 mice; Pg-KO control n = 11; Pg-KO Dsg2-LP n = 12 from 5 mice). Two-way ANOVA with Holm-Šidák post hoc test was performed. (C and F) MEA heatmaps from spontaneously contracting HL-1 cardiomyocytes treated with Dsg2-LP. Pg and Dsg2-levels were reduced by siRNA (Pg-siRNA, Dsg2-siRNA; n.t.-siRNA served as control). Distance between neighboring electrodes: 200 μm. (D, E, G, and H) Corresponding analysis of conduction velocity and SDNN of MEAs as described in C (control n = 5; Trp n = 5; Trp + Dsg2-LP n = 4; Dsg2-LP n = 5), and (F) (n.t.-siRNA n = 5; Pg-siRNA n = 4; Pg-siRNA + Dsg2-LP n = 3; Dsg2-siRNA n = 5; Dsg2-siRNA + Dsg2-LP n = 3). One-way ANOVA with Bonferroni’s post hoc test was performed. (I and J) Western blot analysis shows phosphorylation of Cx43 at serine 368 (pCx43 Ser368) in response to PKC activation by PMA and PKC inhibition by Bim-X or Dsg2-LP in HL-1 (I) and WT mice ventricular cardiac slices (J). Values in top row display mean of densitometric analysis ± SD. Total Cx43 was used as a reference protein. α-Tubulin (α-Tub) served as loading control (n = 4). (K–M) MEAs of Pg-reduced HL-1 cardiomyocytes treated with Bim-X and Dsg2-LP with corresponding analysis of conduction velocity and SDNN (n = 3). *P < 0.05. One-way ANOVA with Bonferroni’s post hoc test.

    Journal: JCI Insight

    Article Title: Stabilization of desmoglein-2 binding rescues arrhythmia in arrhythmogenic cardiomyopathy

    doi: 10.1172/jci.insight.130141

    Figure Lengend Snippet: (A) Representative MEA heatmaps of transversal slices of WT and Pg-KO hearts treated with Dsg2-LP for 1 hour showing the delay of spontaneous excitations relative to a reference electrode (*). Colors correspond to the delay times of excitation given in the scale. Distance between neighboring electrodes: 200 μm. (B) Corresponding analysis of conduction velocity measured in cardiac slices as shown in A (WT control n = 9; WT Dsg2-LP n = 6 from 3 mice; Pg-KO control n = 11; Pg-KO Dsg2-LP n = 12 from 5 mice). Two-way ANOVA with Holm-Šidák post hoc test was performed. (C and F) MEA heatmaps from spontaneously contracting HL-1 cardiomyocytes treated with Dsg2-LP. Pg and Dsg2-levels were reduced by siRNA (Pg-siRNA, Dsg2-siRNA; n.t.-siRNA served as control). Distance between neighboring electrodes: 200 μm. (D, E, G, and H) Corresponding analysis of conduction velocity and SDNN of MEAs as described in C (control n = 5; Trp n = 5; Trp + Dsg2-LP n = 4; Dsg2-LP n = 5), and (F) (n.t.-siRNA n = 5; Pg-siRNA n = 4; Pg-siRNA + Dsg2-LP n = 3; Dsg2-siRNA n = 5; Dsg2-siRNA + Dsg2-LP n = 3). One-way ANOVA with Bonferroni’s post hoc test was performed. (I and J) Western blot analysis shows phosphorylation of Cx43 at serine 368 (pCx43 Ser368) in response to PKC activation by PMA and PKC inhibition by Bim-X or Dsg2-LP in HL-1 (I) and WT mice ventricular cardiac slices (J). Values in top row display mean of densitometric analysis ± SD. Total Cx43 was used as a reference protein. α-Tubulin (α-Tub) served as loading control (n = 4). (K–M) MEAs of Pg-reduced HL-1 cardiomyocytes treated with Bim-X and Dsg2-LP with corresponding analysis of conduction velocity and SDNN (n = 3). *P < 0.05. One-way ANOVA with Bonferroni’s post hoc test.

    Article Snippet: The following primary antibodies were used for Western blot (WB) analysis or immunostaining evaluated by confocal or STED microscopy: monoclonal mouse anti-Dsg1/2 for WB and confocal microscopy (Progen Biotechnik, 61002), polyclonal rabbit anti-Dsg2 for STED (Progen Biotechnik, 610121), monoclonal mouse anti-Pg for WB and confocal microscopy (Progen Biotechnik, 61005), monoclonal mouse anti–α-tubulin for WB (Abcam, ab7291), polyclonal rabbit anti-Cx43 for confocal microscopy and WB (MilliporeSigma, SAB4501175), monoclonal mouse anti-Cx43 for STED (Merck KGaA, MAB3067), phospho-Cx43 Ser368 for WB (Santa Cruz Biotechnology Inc., sc-101660-r), monoclonal mouse anti–N-cadherin (BD Biosciences, 610921), and monoclonal mouse anti-Dp (Progen Biotechnik, 61003).

    Techniques: Western Blot, Activation Assay, Inhibition

    (A) Schematic of atomic force microscope (AFM) force mapping setup. The tip of a cantilever functionalized with the extracellular domains of adhesion molecules via a flexible PEG linker (step 1) is repeatedly extended to (step 2) and retracted from (step 3) a functionalized surface. Interaction of the tip- and surface-bound proteins is detectable by cantilever deflection. (B and C) Analysis of unbinding force and binding frequency of cell-free AFM measurements with Dsg2- or N-Cad–coated tips probed on mica sheets with respective coatings. Every dot in the right panel represents the mean value of 1 independent experiment, bars indicate mean ± SD, and black lines connect paired values. n = 6 for Dsg2; n = 5 for N-Cad. n.s., P ≥ 0.05, *P < 0.05. Two-tailed paired Student’s t test.

    Journal: JCI Insight

    Article Title: Stabilization of desmoglein-2 binding rescues arrhythmia in arrhythmogenic cardiomyopathy

    doi: 10.1172/jci.insight.130141

    Figure Lengend Snippet: (A) Schematic of atomic force microscope (AFM) force mapping setup. The tip of a cantilever functionalized with the extracellular domains of adhesion molecules via a flexible PEG linker (step 1) is repeatedly extended to (step 2) and retracted from (step 3) a functionalized surface. Interaction of the tip- and surface-bound proteins is detectable by cantilever deflection. (B and C) Analysis of unbinding force and binding frequency of cell-free AFM measurements with Dsg2- or N-Cad–coated tips probed on mica sheets with respective coatings. Every dot in the right panel represents the mean value of 1 independent experiment, bars indicate mean ± SD, and black lines connect paired values. n = 6 for Dsg2; n = 5 for N-Cad. n.s., P ≥ 0.05, *P < 0.05. Two-tailed paired Student’s t test.

    Article Snippet: The following primary antibodies were used for Western blot (WB) analysis or immunostaining evaluated by confocal or STED microscopy: monoclonal mouse anti-Dsg1/2 for WB and confocal microscopy (Progen Biotechnik, 61002), polyclonal rabbit anti-Dsg2 for STED (Progen Biotechnik, 610121), monoclonal mouse anti-Pg for WB and confocal microscopy (Progen Biotechnik, 61005), monoclonal mouse anti–α-tubulin for WB (Abcam, ab7291), polyclonal rabbit anti-Cx43 for confocal microscopy and WB (MilliporeSigma, SAB4501175), monoclonal mouse anti-Cx43 for STED (Merck KGaA, MAB3067), phospho-Cx43 Ser368 for WB (Santa Cruz Biotechnology Inc., sc-101660-r), monoclonal mouse anti–N-cadherin (BD Biosciences, 610921), and monoclonal mouse anti-Dp (Progen Biotechnik, 61003).

    Techniques: Microscopy, Binding Assay, Two Tailed Test

    Representative binding event and topography images of AFM force mapping performed with a Dsg2-coated tip on HL-1 cardiomyocytes (A) or cardiomyocytes isolated from Dsg2-KO mice or WT littermates (D, G, and J) treated with Dsg2-LP 30–90 minutes. Cyan line indicates cell-cell border. Every white pixel represents 1 binding event at the respective location. (A) n = 5 independent experiments, scale bar: 1 μm. (D) n = 7 mice, (G) n = 6 mice per phenotype, and (J) n = 6 mice; scale bar: 2 μm. Unbinding forces (B, E, H, and K) and binding frequency (C, F, I, and L) of Dsg2-mediated binding events corresponding to respective panels on the left. Every dot represents the mean value of 1 independent experiment, bars indicate mean ± SD, and black lines connect paired values. *P < 0.05. Two-tailed paired Student’s t test was performed.

    Journal: JCI Insight

    Article Title: Stabilization of desmoglein-2 binding rescues arrhythmia in arrhythmogenic cardiomyopathy

    doi: 10.1172/jci.insight.130141

    Figure Lengend Snippet: Representative binding event and topography images of AFM force mapping performed with a Dsg2-coated tip on HL-1 cardiomyocytes (A) or cardiomyocytes isolated from Dsg2-KO mice or WT littermates (D, G, and J) treated with Dsg2-LP 30–90 minutes. Cyan line indicates cell-cell border. Every white pixel represents 1 binding event at the respective location. (A) n = 5 independent experiments, scale bar: 1 μm. (D) n = 7 mice, (G) n = 6 mice per phenotype, and (J) n = 6 mice; scale bar: 2 μm. Unbinding forces (B, E, H, and K) and binding frequency (C, F, I, and L) of Dsg2-mediated binding events corresponding to respective panels on the left. Every dot represents the mean value of 1 independent experiment, bars indicate mean ± SD, and black lines connect paired values. *P < 0.05. Two-tailed paired Student’s t test was performed.

    Article Snippet: The following primary antibodies were used for Western blot (WB) analysis or immunostaining evaluated by confocal or STED microscopy: monoclonal mouse anti-Dsg1/2 for WB and confocal microscopy (Progen Biotechnik, 61002), polyclonal rabbit anti-Dsg2 for STED (Progen Biotechnik, 610121), monoclonal mouse anti-Pg for WB and confocal microscopy (Progen Biotechnik, 61005), monoclonal mouse anti–α-tubulin for WB (Abcam, ab7291), polyclonal rabbit anti-Cx43 for confocal microscopy and WB (MilliporeSigma, SAB4501175), monoclonal mouse anti-Cx43 for STED (Merck KGaA, MAB3067), phospho-Cx43 Ser368 for WB (Santa Cruz Biotechnology Inc., sc-101660-r), monoclonal mouse anti–N-cadherin (BD Biosciences, 610921), and monoclonal mouse anti-Dp (Progen Biotechnik, 61003).

    Techniques: Binding Assay, Isolation, Two Tailed Test

    (A) Membrane-impermeable cross-linking with sulfo-EGS performed after 24-hour treatment with Dsg2-LP in HL-1 cells. Western blot analyses were done for Dsg2 and N-Cad. α-Tubulin (α-Tub) served as loading control. Representative image of 4 repeated experiments. (B) Densitometric quantification of Dsg2 and N-Cad bands showing the ratio of cross-linked proteins to total protein. n = 4. *P < 0.05. Two-tailed paired Student’s t test with 95% confidence levels was performed.

    Journal: JCI Insight

    Article Title: Stabilization of desmoglein-2 binding rescues arrhythmia in arrhythmogenic cardiomyopathy

    doi: 10.1172/jci.insight.130141

    Figure Lengend Snippet: (A) Membrane-impermeable cross-linking with sulfo-EGS performed after 24-hour treatment with Dsg2-LP in HL-1 cells. Western blot analyses were done for Dsg2 and N-Cad. α-Tubulin (α-Tub) served as loading control. Representative image of 4 repeated experiments. (B) Densitometric quantification of Dsg2 and N-Cad bands showing the ratio of cross-linked proteins to total protein. n = 4. *P < 0.05. Two-tailed paired Student’s t test with 95% confidence levels was performed.

    Article Snippet: The following primary antibodies were used for Western blot (WB) analysis or immunostaining evaluated by confocal or STED microscopy: monoclonal mouse anti-Dsg1/2 for WB and confocal microscopy (Progen Biotechnik, 61002), polyclonal rabbit anti-Dsg2 for STED (Progen Biotechnik, 610121), monoclonal mouse anti-Pg for WB and confocal microscopy (Progen Biotechnik, 61005), monoclonal mouse anti–α-tubulin for WB (Abcam, ab7291), polyclonal rabbit anti-Cx43 for confocal microscopy and WB (MilliporeSigma, SAB4501175), monoclonal mouse anti-Cx43 for STED (Merck KGaA, MAB3067), phospho-Cx43 Ser368 for WB (Santa Cruz Biotechnology Inc., sc-101660-r), monoclonal mouse anti–N-cadherin (BD Biosciences, 610921), and monoclonal mouse anti-Dp (Progen Biotechnik, 61003).

    Techniques: Western Blot, Two Tailed Test