nucleosome structures  (Worthington Biochemical)


Bioz Verified Symbol Worthington Biochemical is a verified supplier
Bioz Manufacturer Symbol Worthington Biochemical manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 85
    Name:
    Neonatal Cardiomyocyte Isolation System
    Description:
    Kit for performing five separate tissue dissociations each containing up to twelve hearts Contains single use vials of purified collagenase and trypsin CMF HBSS Leibovitz L 15 media and Falcon cell strainers along with a detailed protocol The kit is use tested by Worthington to assure performance
    Catalog Number:
    lk003300
    Price:
    256
    Size:
    1 kt
    Cas Number:
    see components
    Buy from Supplier


    Structured Review

    Worthington Biochemical nucleosome structures
    Remodeling of <t>dinucleosomes</t> by ISWI complexes stimulates histone methylation by SET domain proteins. (A) The SET domain of SET7 binds histones, but not <t>nucleosomes.</t> GST pulldown experiments were conducted with immobilized GST-SET7 polypeptides (residues
    Kit for performing five separate tissue dissociations each containing up to twelve hearts Contains single use vials of purified collagenase and trypsin CMF HBSS Leibovitz L 15 media and Falcon cell strainers along with a detailed protocol The kit is use tested by Worthington to assure performance
    https://www.bioz.com/result/nucleosome structures/product/Worthington Biochemical
    Average 85 stars, based on 209 article reviews
    Price from $9.99 to $1999.99
    nucleosome structures - by Bioz Stars, 2020-08
    85/100 stars

    Images

    1) Product Images from "SET Domains of Histone Methyltransferases Recognize ISWI-Remodeled Nucleosomal Species ▿"

    Article Title: SET Domains of Histone Methyltransferases Recognize ISWI-Remodeled Nucleosomal Species ▿

    Journal: Molecular and Cellular Biology

    doi: 10.1128/MCB.00775-09

    Remodeling of dinucleosomes by ISWI complexes stimulates histone methylation by SET domain proteins. (A) The SET domain of SET7 binds histones, but not nucleosomes. GST pulldown experiments were conducted with immobilized GST-SET7 polypeptides (residues
    Figure Legend Snippet: Remodeling of dinucleosomes by ISWI complexes stimulates histone methylation by SET domain proteins. (A) The SET domain of SET7 binds histones, but not nucleosomes. GST pulldown experiments were conducted with immobilized GST-SET7 polypeptides (residues

    Techniques Used: Methylation

    Analysis of ISW2 and SWI/SNF remodeled dinucleosomes. 32 P-end-labeled dinucleosomes were remodeled by ISW2 (A and B) or SWI/SNF (C and D) as described in the legend for Fig. . Remodeling was terminated by apyrase treatment, and nucleosomes
    Figure Legend Snippet: Analysis of ISW2 and SWI/SNF remodeled dinucleosomes. 32 P-end-labeled dinucleosomes were remodeled by ISW2 (A and B) or SWI/SNF (C and D) as described in the legend for Fig. . Remodeling was terminated by apyrase treatment, and nucleosomes

    Techniques Used: Labeling

    The SET domain of trithorax binds core histones and altered nucleosomal structures but not intact nucleosomes. (A) Schematic of the domain structure of trithorax. Positions of highly conserved blocks of homology with methyltransferases, a C-terminal cysteine-rich
    Figure Legend Snippet: The SET domain of trithorax binds core histones and altered nucleosomal structures but not intact nucleosomes. (A) Schematic of the domain structure of trithorax. Positions of highly conserved blocks of homology with methyltransferases, a C-terminal cysteine-rich

    Techniques Used:

    The SET domain of ALL1 does not bind remodeled mononucleosomes. (A, top) Analysis of remodeled mononucleosomes by native PAGE. Remodeling assay mixtures (50 μl) contained 1.2 μg of nucleosomes and 2.5 ng of ISWI or 5 ng of Swi-Snf complexes
    Figure Legend Snippet: The SET domain of ALL1 does not bind remodeled mononucleosomes. (A, top) Analysis of remodeled mononucleosomes by native PAGE. Remodeling assay mixtures (50 μl) contained 1.2 μg of nucleosomes and 2.5 ng of ISWI or 5 ng of Swi-Snf complexes

    Techniques Used: Clear Native PAGE

    The SET domain of ALL1 binds dinucleosomes remodeled by the ISWI class of chromatin remodeling enzymes. (A) Example of dinucleosome assembly. Dinucleosomes were reconstituted onto DNA containing two 601 minimal nucleosome positioning sequences (the orientation
    Figure Legend Snippet: The SET domain of ALL1 binds dinucleosomes remodeled by the ISWI class of chromatin remodeling enzymes. (A) Example of dinucleosome assembly. Dinucleosomes were reconstituted onto DNA containing two 601 minimal nucleosome positioning sequences (the orientation

    Techniques Used:

    2) Product Images from "Kinetics of rate-dependent shortening of action potential duration in guinea-pig ventricle; effects of IK1 and IKr blockade"

    Article Title: Kinetics of rate-dependent shortening of action potential duration in guinea-pig ventricle; effects of IK1 and IKr blockade

    Journal: British Journal of Pharmacology

    doi: 10.1038/sj.bjp.0702443

    The time course of APD 90 shortening following a cycle length change from 450 ms to 300 ms in control for a Langendorff-perfused ventricular preparation, showing the two types of exponential fit. Zero on the time axis is the time at which the cycle length was changed. In the main plot, the data was fitted with a mono-exponential (solid line) and, in the inset plot, the same data was fitted with a bi-exponential. The first 50 s of shortening are clearly better fitted with a bi-exponential. Of nine experiments, all could be fit with a mono-exponential, although in five experiments, the first 30–50 s of data were better fit with a bi-exponential.
    Figure Legend Snippet: The time course of APD 90 shortening following a cycle length change from 450 ms to 300 ms in control for a Langendorff-perfused ventricular preparation, showing the two types of exponential fit. Zero on the time axis is the time at which the cycle length was changed. In the main plot, the data was fitted with a mono-exponential (solid line) and, in the inset plot, the same data was fitted with a bi-exponential. The first 50 s of shortening are clearly better fitted with a bi-exponential. Of nine experiments, all could be fit with a mono-exponential, although in five experiments, the first 30–50 s of data were better fit with a bi-exponential.

    Techniques Used: Mass Spectrometry

    3) Product Images from "Nucleosomes around a mismatched base pair are excluded via an Msh2-dependent reaction with the aid of SNF2 family ATPase Smarcad1"

    Article Title: Nucleosomes around a mismatched base pair are excluded via an Msh2-dependent reaction with the aid of SNF2 family ATPase Smarcad1

    Journal: Genes & Development

    doi: 10.1101/gad.310995.117

    Nucleosomes are excluded from a > 1-kb region surrounding a mismatch. ( A ) The DNA substrate used in this study. The 3011-base-pair (bp) DNA carries an A:T base pair (pMM1 homo ) or an A:C mispair (pMM1 AC ) at position 1. Positions of restriction enzyme sites used in this study, the site of biotin modification, and amplicons for quantitative PCR (qPCR) (P1: 2950–61, P2: 253–383, P3: 476–602, P4: 728–860, P5: 1498–1628, P6: 2266–2397, and P7: 2413–2537) are indicated. ( B ) Supercoiling assay in NPE. Covalently closed pMM1 homo (lanes 2 – 8 ) or pMM1 AC (lanes 9 – 15 ) was incubated in NPE and sampled at the indicated times. (Lane 1 ) Supercoiled pMM1 homo purified from Escherichia coli was used as a size standard. (oc/r) Open circular or relaxed DNA; (sc) supercoiled DNA. ( C ) pMM1 homo (lanes 1 – 4 ) or pMM1 AC (lanes 5 – 8 ) was incubated in NPE for 60 min and digested by micrococcal nuclease (MNase). DNA samples stained with SYBR Gold ( top ) and Southern blotting with the PvuII–PvuII probe ( middle ) and the DraI–DraI probe ( bottom ) are shown. ( D – F ) The MNase assay described in C was repeated in the presence of a control plasmid (pControl), and undigested DNA was quantified by qPCR. The amount of DNA relative to the input ( D ) and normalized to pControl ( E ) and pMM1 homo ( F ) is presented. Mean ± one standard deviation (SD) is shown. n = 3.
    Figure Legend Snippet: Nucleosomes are excluded from a > 1-kb region surrounding a mismatch. ( A ) The DNA substrate used in this study. The 3011-base-pair (bp) DNA carries an A:T base pair (pMM1 homo ) or an A:C mispair (pMM1 AC ) at position 1. Positions of restriction enzyme sites used in this study, the site of biotin modification, and amplicons for quantitative PCR (qPCR) (P1: 2950–61, P2: 253–383, P3: 476–602, P4: 728–860, P5: 1498–1628, P6: 2266–2397, and P7: 2413–2537) are indicated. ( B ) Supercoiling assay in NPE. Covalently closed pMM1 homo (lanes 2 – 8 ) or pMM1 AC (lanes 9 – 15 ) was incubated in NPE and sampled at the indicated times. (Lane 1 ) Supercoiled pMM1 homo purified from Escherichia coli was used as a size standard. (oc/r) Open circular or relaxed DNA; (sc) supercoiled DNA. ( C ) pMM1 homo (lanes 1 – 4 ) or pMM1 AC (lanes 5 – 8 ) was incubated in NPE for 60 min and digested by micrococcal nuclease (MNase). DNA samples stained with SYBR Gold ( top ) and Southern blotting with the PvuII–PvuII probe ( middle ) and the DraI–DraI probe ( bottom ) are shown. ( D – F ) The MNase assay described in C was repeated in the presence of a control plasmid (pControl), and undigested DNA was quantified by qPCR. The amount of DNA relative to the input ( D ) and normalized to pControl ( E ) and pMM1 homo ( F ) is presented. Mean ± one standard deviation (SD) is shown. n = 3.

    Techniques Used: Modification, Real-time Polymerase Chain Reaction, Incubation, Purification, Staining, Southern Blot, Plasmid Preparation, Standard Deviation

    4) Product Images from "Pivotal role of cardiac lineage protein-1 (CLP-1) in transcriptional elongation factor P-TEFb complex formation in cardiac hypertrophy"

    Article Title: Pivotal role of cardiac lineage protein-1 (CLP-1) in transcriptional elongation factor P-TEFb complex formation in cardiac hypertrophy

    Journal:

    doi: 10.1016/j.cardiores.2007.03.019

    Immunocytochemical co-localization of CLP-1, cyclin T and cdk9 in isolated 2 day post natal rat cardiomyocytes. A–C: (A) CLP-1 is in the nucleus of MF20-positive cardiomyocytes; (B) nuclear co-localization of CLP-1 and cyclin T; (C) control (no
    Figure Legend Snippet: Immunocytochemical co-localization of CLP-1, cyclin T and cdk9 in isolated 2 day post natal rat cardiomyocytes. A–C: (A) CLP-1 is in the nucleus of MF20-positive cardiomyocytes; (B) nuclear co-localization of CLP-1 and cyclin T; (C) control (no

    Techniques Used: Isolation

    A. Stretched cardiomyocytes express markers of hypertrophy. Northern blot showing increased ANF (atrial natriuretic factor) mRNA levels in stretched cardiomyocytes. GAPDH is loading control. B. Phosphorylation of STAT 3 increases with continued stretch
    Figure Legend Snippet: A. Stretched cardiomyocytes express markers of hypertrophy. Northern blot showing increased ANF (atrial natriuretic factor) mRNA levels in stretched cardiomyocytes. GAPDH is loading control. B. Phosphorylation of STAT 3 increases with continued stretch

    Techniques Used: Northern Blot

    CLP-1 dissociates from P-TEFb in cardiomyocytes treated with hypertrophic agents. A. Immunoprecipitated cyclin T1 shows less associated CLP-1 in endothelin-1-treated cardiomyocytes than untreated cells (control). B. Immunoprecipitated cyclin T1 shows
    Figure Legend Snippet: CLP-1 dissociates from P-TEFb in cardiomyocytes treated with hypertrophic agents. A. Immunoprecipitated cyclin T1 shows less associated CLP-1 in endothelin-1-treated cardiomyocytes than untreated cells (control). B. Immunoprecipitated cyclin T1 shows

    Techniques Used: Immunoprecipitation

    5) Product Images from "An siRNA Screen Identifies the U2 snRNP Spliceosome as a Host Restriction Factor for Recombinant Adeno-associated Viruses"

    Article Title: An siRNA Screen Identifies the U2 snRNP Spliceosome as a Host Restriction Factor for Recombinant Adeno-associated Viruses

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1005082

    Meayamycin B increases AAV vector transduction of clinically relevant cell types. (A) Primary mouse islets were infected with AAV8 CMV-GFP in the presence or absence of 2 nM meayamycin B, and GFP expression was monitored for three days. (B) Primary human islets were treated with AAV2 or AAV9 CMV-Luc vectors for 7 hours and then treated with 0, 2, 5 or 10 nM meayamycin B. Luciferase expression was analyzed 48 hours p.i. (C) Neonatal rat cardiomyocytes were infected with AAV2 CMV-Luc or scAAV9 CMV-GFP vectors and treated with meayamycin B, 3 hours p.i. Luciferase activity was measured 3 days p.i., while GFP expression was monitored at 5 days p.i. (D) Porcine hepatocytes were infected with AAV2 or AAV9 CMV-Luc vectors for 8 hours, virus was then removed and cells were treated with 0, 2, or 20 nM meayamycin B. Cells were harvested 48 hours p.i. for the luciferase assay. In A-D, an MOI of 10 4 was used.
    Figure Legend Snippet: Meayamycin B increases AAV vector transduction of clinically relevant cell types. (A) Primary mouse islets were infected with AAV8 CMV-GFP in the presence or absence of 2 nM meayamycin B, and GFP expression was monitored for three days. (B) Primary human islets were treated with AAV2 or AAV9 CMV-Luc vectors for 7 hours and then treated with 0, 2, 5 or 10 nM meayamycin B. Luciferase expression was analyzed 48 hours p.i. (C) Neonatal rat cardiomyocytes were infected with AAV2 CMV-Luc or scAAV9 CMV-GFP vectors and treated with meayamycin B, 3 hours p.i. Luciferase activity was measured 3 days p.i., while GFP expression was monitored at 5 days p.i. (D) Porcine hepatocytes were infected with AAV2 or AAV9 CMV-Luc vectors for 8 hours, virus was then removed and cells were treated with 0, 2, or 20 nM meayamycin B. Cells were harvested 48 hours p.i. for the luciferase assay. In A-D, an MOI of 10 4 was used.

    Techniques Used: Plasmid Preparation, Transduction, Infection, Expressing, Luciferase, Activity Assay

    6) Product Images from "Single Cell Transcriptomics Reconstructs Fate Conversion from Fibroblast to Cardiomyocyte"

    Article Title: Single Cell Transcriptomics Reconstructs Fate Conversion from Fibroblast to Cardiomyocyte

    Journal: Nature

    doi: 10.1038/nature24454

    Heterogeneity of our isolated CFs (Thy1+ non-immune non-myocyte cardiac cells) and stepwise suppression of non-cardiomyocyte lineages during iCM reprogramming Related to . ( a-c ) Limited transcriptome change by retrovirus transduction. To determine whether introduction of viruses could influence cellular identities of CF, molecular features of the uninfected and DsRed-transduced cells were compared and only 25 genes were differentially expressed (ANOVA p value
    Figure Legend Snippet: Heterogeneity of our isolated CFs (Thy1+ non-immune non-myocyte cardiac cells) and stepwise suppression of non-cardiomyocyte lineages during iCM reprogramming Related to . ( a-c ) Limited transcriptome change by retrovirus transduction. To determine whether introduction of viruses could influence cellular identities of CF, molecular features of the uninfected and DsRed-transduced cells were compared and only 25 genes were differentially expressed (ANOVA p value

    Techniques Used: Isolation, Transduction

    Heterogeneity of CF and stepwise suppression of non-cardiomyocyte lineages during iCM induction ( a-b ) HC (a) and PCA (b) of control CFs with representative gene expression and GO analysis of the five identified gene clusters. ( c ) HC calculated with control CFs (a) applied to M+G+T-transduced cells with representative gene expression. ( d-f ) 40× ICC images (d,e) with quantifications (f) of Thy1 and SM22α co-stained with αMHC-GFP during reprogramming. n=20 images, scale bar=100 μm, error bars indicate SEM.
    Figure Legend Snippet: Heterogeneity of CF and stepwise suppression of non-cardiomyocyte lineages during iCM induction ( a-b ) HC (a) and PCA (b) of control CFs with representative gene expression and GO analysis of the five identified gene clusters. ( c ) HC calculated with control CFs (a) applied to M+G+T-transduced cells with representative gene expression. ( d-f ) 40× ICC images (d,e) with quantifications (f) of Thy1 and SM22α co-stained with αMHC-GFP during reprogramming. n=20 images, scale bar=100 μm, error bars indicate SEM.

    Techniques Used: Expressing, Immunocytochemistry, Staining

    7) Product Images from "Muscle ring finger protein-1 inhibits PKC? activation and prevents cardiomyocyte hypertrophy"

    Article Title: Muscle ring finger protein-1 inhibits PKC? activation and prevents cardiomyocyte hypertrophy

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.200402033

    MURF1 colocalizes with RACK1 in cultured cardiac myocytes.  Rat cardiac myocytes were infected with recombinant adenovirus Ad.GFP (A and B) or Ad.MURF1 (C and D) for 24 h in serum free medium followed by induction with PE (B and D) for 48 h. Immunostaining was done with anti-Myc and anti-RACK1 antibodies. This was followed by secondary antibody incubation with anti–rabbit Alexa 568 (red) and anti–mouse AMCA (blue). The green color represents GFP expression.
    Figure Legend Snippet: MURF1 colocalizes with RACK1 in cultured cardiac myocytes. Rat cardiac myocytes were infected with recombinant adenovirus Ad.GFP (A and B) or Ad.MURF1 (C and D) for 24 h in serum free medium followed by induction with PE (B and D) for 48 h. Immunostaining was done with anti-Myc and anti-RACK1 antibodies. This was followed by secondary antibody incubation with anti–rabbit Alexa 568 (red) and anti–mouse AMCA (blue). The green color represents GFP expression.

    Techniques Used: Cell Culture, Infection, Recombinant, Immunostaining, Incubation, Expressing

    8) Product Images from "Three-Dimensional Culture Alters Primary Cardiac Cell Phenotype"

    Article Title: Three-Dimensional Culture Alters Primary Cardiac Cell Phenotype

    Journal: Tissue Engineering. Part A

    doi: 10.1089/ten.tea.2009.0458

    Assessment of homogeneity between two-dimensional (2D) and three-dimensional (3D) cultures. Samples were collected on day 1 or 6 of culture as indicated and assessed for bulk differences in culture composition, hypertrophy, and metabolic function. ( A ) Cell attachment efficiency was calculated as the number of cells attached to tissue culture polystyrene (TCPS) surfaces after 24 h divided by the total number of viable cells in the original inoculum. The proportion of attached cardiomyocytes (CMs), which are positive for myosin heavy chain (MyHC) immunostaining, per total adherent cells was also calculated. ( B ) On day 6, MyHC and filamentous actin (f-Actin) levels, which are indicative of CM hypertrophic status, were determined in fixed samples by fluorescence in situ quantitation assay and are presented as fluorescence ratios normalized to DNA. Total protein content, which is a key indicator of tissue hypertrophy, was measured in culture homogenates and normalized to DNA. Units presented are μg protein per 10 ng DNA. ( C ) Intermediary metabolic enzyme activities in day 6 cell extracts calculated as units of enzyme activity per mg of total protein per minute. No significant differences were found in ( A ), ( B ), or ( C ). Data are mean values ± standard deviation for n = 6 samples.
    Figure Legend Snippet: Assessment of homogeneity between two-dimensional (2D) and three-dimensional (3D) cultures. Samples were collected on day 1 or 6 of culture as indicated and assessed for bulk differences in culture composition, hypertrophy, and metabolic function. ( A ) Cell attachment efficiency was calculated as the number of cells attached to tissue culture polystyrene (TCPS) surfaces after 24 h divided by the total number of viable cells in the original inoculum. The proportion of attached cardiomyocytes (CMs), which are positive for myosin heavy chain (MyHC) immunostaining, per total adherent cells was also calculated. ( B ) On day 6, MyHC and filamentous actin (f-Actin) levels, which are indicative of CM hypertrophic status, were determined in fixed samples by fluorescence in situ quantitation assay and are presented as fluorescence ratios normalized to DNA. Total protein content, which is a key indicator of tissue hypertrophy, was measured in culture homogenates and normalized to DNA. Units presented are μg protein per 10 ng DNA. ( C ) Intermediary metabolic enzyme activities in day 6 cell extracts calculated as units of enzyme activity per mg of total protein per minute. No significant differences were found in ( A ), ( B ), or ( C ). Data are mean values ± standard deviation for n = 6 samples.

    Techniques Used: Cell Attachment Assay, Immunostaining, Fluorescence, In Situ, Quantitation Assay, Activity Assay, Standard Deviation

    Distribution of cells in 3D aggregates. ( A ) Phase contrast image of a standard 2D culture on TCPS culture wells. Arrows indicate patches of elongated cells (lower arrow) indicative of cardiac muscle cells interspersed with nonmuscle cells (upper arrow). ( B ) Phase contrast image of 3D aggregates initiated on TCPS support beads. Arrow indicates typical 3D cell mass. Inset is a lower magnification view of a multi-cell/multi-bead aggregate. ( C ) Two-dimensional culture stained for MyHC to indicate CMs and DNA. Similar to ( A ), patches of cardiac muscle cells are interspersed with nonmuscle cells (arrows) in a mosaic pattern. ( D ) Three-dimensional culture stained for MyHC to indicate CMs and DNA. The exterior-most cells do not contain MyHC (arrows indicate nuclei of these cells). ( E ) Three-dimensional culture stained for vimentin and DNA, verifying the presence of mesenchymal/endothelial cells (ECs) on the aggregate surface (arrows); this staining is consistent with an EC layer. ( F ) Two-dimensional culture stained for platelet/endothelial cell adhesion molecule (PECAM) (CD31) and DNA showing an island of PECAM-positive ECs amidst PECAM-negative cells (arrow). ( G ) Three-dimensional culture stained for PECAM showing that exterior-most nuclei are within cells positive for PECAM (arrow), a marker for ECs in contact with each other. ( H ) Scanning electron micrograph (SEM) of a 2D culture. Lower arrow indicates an elongated, binucleate cell, which is consistent with a CM. Upper arrow indicates a mononucleated cell with a more spread morphology. Nuceoli are readily apparent. ( I ) SEM of a 3D culture surface showing only cells with an endothelial appearance are present (arrows). ( J ) SEM of the interior aspect of a rat heart lumen showing the configuration of the ECs lining the organ lumen. Bar = 50 μm except for inset figure in ( B .
    Figure Legend Snippet: Distribution of cells in 3D aggregates. ( A ) Phase contrast image of a standard 2D culture on TCPS culture wells. Arrows indicate patches of elongated cells (lower arrow) indicative of cardiac muscle cells interspersed with nonmuscle cells (upper arrow). ( B ) Phase contrast image of 3D aggregates initiated on TCPS support beads. Arrow indicates typical 3D cell mass. Inset is a lower magnification view of a multi-cell/multi-bead aggregate. ( C ) Two-dimensional culture stained for MyHC to indicate CMs and DNA. Similar to ( A ), patches of cardiac muscle cells are interspersed with nonmuscle cells (arrows) in a mosaic pattern. ( D ) Three-dimensional culture stained for MyHC to indicate CMs and DNA. The exterior-most cells do not contain MyHC (arrows indicate nuclei of these cells). ( E ) Three-dimensional culture stained for vimentin and DNA, verifying the presence of mesenchymal/endothelial cells (ECs) on the aggregate surface (arrows); this staining is consistent with an EC layer. ( F ) Two-dimensional culture stained for platelet/endothelial cell adhesion molecule (PECAM) (CD31) and DNA showing an island of PECAM-positive ECs amidst PECAM-negative cells (arrow). ( G ) Three-dimensional culture stained for PECAM showing that exterior-most nuclei are within cells positive for PECAM (arrow), a marker for ECs in contact with each other. ( H ) Scanning electron micrograph (SEM) of a 2D culture. Lower arrow indicates an elongated, binucleate cell, which is consistent with a CM. Upper arrow indicates a mononucleated cell with a more spread morphology. Nuceoli are readily apparent. ( I ) SEM of a 3D culture surface showing only cells with an endothelial appearance are present (arrows). ( J ) SEM of the interior aspect of a rat heart lumen showing the configuration of the ECs lining the organ lumen. Bar = 50 μm except for inset figure in ( B .

    Techniques Used: Staining, Marker

    9) Product Images from "NO triggers RGS4 degradation to coordinate angiogenesis and cardiomyocyte growth"

    Article Title: NO triggers RGS4 degradation to coordinate angiogenesis and cardiomyocyte growth

    Journal: The Journal of Clinical Investigation

    doi: 10.1172/JCI65112

    NO mediates cardiomyocyte hypertrophy in culture.
    Figure Legend Snippet: NO mediates cardiomyocyte hypertrophy in culture.

    Techniques Used:

    Endothelium-to-cardiomyocyte communication paradigm.
    Figure Legend Snippet: Endothelium-to-cardiomyocyte communication paradigm.

    Techniques Used:

    10) Product Images from "Mitoregulin: A lncRNA-Encoded Microprotein that Supports Mitochondrial Supercomplexes and Respiratory Efficiency"

    Article Title: Mitoregulin: A lncRNA-Encoded Microprotein that Supports Mitochondrial Supercomplexes and Respiratory Efficiency

    Journal: Cell reports

    doi: 10.1016/j.celrep.2018.06.002

    The LINC00116 -Derived Microprotein Mitoregulin Localizes to Inner Mitochondrial Membranes and Binds Cardiolipin (A and B) Wild-type (A) and GFP-tagged (B) human Mtln were expressed in neonatal rat cardiomyocytes, and co-localization with MitoTracker red was evaluated. Representative photomicrographs are shown. Scale bars, 10 μm. (C) Mitochondrial pellets were isolated from wild-type (WT) or Mtln-knockout (KO) C2C12 myoblast cells, and western blot was performed on various fractions. (D) Mitochondrial pellets harvested from WT or Mtln-KO skeletal muscle tissues were treated with increasing digitonin concentrations to release OMMs, and pellet and supernatant fraction fractions were subjected to western blot analysis. Cox4 and Vdac1 are known IMM and OMM proteins, respectively. Gapdh is a cytosolic protein known to associate with mitochondria in some cases. (E) Mitochondrial pellets harvested from WT skeletal muscle tissues were resuspended in isotonic, hypotonic, or isotonic plus triton buffers in the absence or presence of proteinase K and subjected to western blot analysis. Proteins with known localization to various mitochondrial compartments (e.g., matrix, IMM, and intermembrane space [IMS]) were evaluated as controls. (F) Western blot analysis performed on WT and Mtln-KO cardiac tissue lysates subjected to pull-down assay using cardiolipin (CL)-coated or control beads. Subunit c, a known cardiolipin-binding protein, serves as the positive control. (G) Lipid-strip binding assay performed using synthetic Mtln protein followed by anti-Mtln immunoblot.
    Figure Legend Snippet: The LINC00116 -Derived Microprotein Mitoregulin Localizes to Inner Mitochondrial Membranes and Binds Cardiolipin (A and B) Wild-type (A) and GFP-tagged (B) human Mtln were expressed in neonatal rat cardiomyocytes, and co-localization with MitoTracker red was evaluated. Representative photomicrographs are shown. Scale bars, 10 μm. (C) Mitochondrial pellets were isolated from wild-type (WT) or Mtln-knockout (KO) C2C12 myoblast cells, and western blot was performed on various fractions. (D) Mitochondrial pellets harvested from WT or Mtln-KO skeletal muscle tissues were treated with increasing digitonin concentrations to release OMMs, and pellet and supernatant fraction fractions were subjected to western blot analysis. Cox4 and Vdac1 are known IMM and OMM proteins, respectively. Gapdh is a cytosolic protein known to associate with mitochondria in some cases. (E) Mitochondrial pellets harvested from WT skeletal muscle tissues were resuspended in isotonic, hypotonic, or isotonic plus triton buffers in the absence or presence of proteinase K and subjected to western blot analysis. Proteins with known localization to various mitochondrial compartments (e.g., matrix, IMM, and intermembrane space [IMS]) were evaluated as controls. (F) Western blot analysis performed on WT and Mtln-KO cardiac tissue lysates subjected to pull-down assay using cardiolipin (CL)-coated or control beads. Subunit c, a known cardiolipin-binding protein, serves as the positive control. (G) Lipid-strip binding assay performed using synthetic Mtln protein followed by anti-Mtln immunoblot.

    Techniques Used: Derivative Assay, Isolation, Knock-Out, Western Blot, Pull Down Assay, Binding Assay, Positive Control, Stripping Membranes

    11) Product Images from "Control of histone H3 phosphorylation by CaMKIIδ in response to haemodynamic cardiac stress"

    Article Title: Control of histone H3 phosphorylation by CaMKIIδ in response to haemodynamic cardiac stress

    Journal: The Journal of Pathology

    doi: 10.1002/path.4489

    14–3–3 recruitment to chromatin is important for transcriptional elongation of GATA-4 and of fetal cardiac genes. (A) ChIP–qPCR performed from mouse heart tissues showing 14–3–3 binding at the ANP and β -MHC promoters in CaMKII δ -WT or CaMKII δ -KO mice 21 days after sham operation or TAC. qPCR reactions were also performed at GAPDH and α-actin promoters that are not responsive to pressure overload hypertrophy: results are expressed as percentage input over signals with IgG antibody and represent average ± SD ( n = 3); p values from Student's t test are indicated. (B) Immunoblotting of cardiomyocytes expressing reduced 14–3–3 (si14–3–3) or normal 14–3–3 (siControl). (C) ChIP–qPCR, showing p-RNAPII S2 at the promoter and internal exon of ANP , β-MHC , GATA-4 , GAPDH and actin in primary neonatal rat cardiomyocytes with siCt or si14–3–3 and stimulated with phenylephrine (PE) for 24 h. (D) ChIP–qPCR assay from mouse heart chromatin, showing p-RNAPII S2 at the ANP , β-MHC , actin and GAPDH exonic region in CaMKII δ -WT or CaMKII δ -KO animals, 21 days after sham operation or TAC. (E) ChIP–qPCR assay, showing p-RNAPII S2 at the ANP , β-MHC , actin and GAPDH promoter regions in CaMKII δ -WT or CaMKII δ -KO animals, 21 days after sham operation or TAC: results are expressed as percentage input over signals with IgG antibody, and represent average ± SD ( n = 3); p values from Student's t -test are indicated
    Figure Legend Snippet: 14–3–3 recruitment to chromatin is important for transcriptional elongation of GATA-4 and of fetal cardiac genes. (A) ChIP–qPCR performed from mouse heart tissues showing 14–3–3 binding at the ANP and β -MHC promoters in CaMKII δ -WT or CaMKII δ -KO mice 21 days after sham operation or TAC. qPCR reactions were also performed at GAPDH and α-actin promoters that are not responsive to pressure overload hypertrophy: results are expressed as percentage input over signals with IgG antibody and represent average ± SD ( n = 3); p values from Student's t test are indicated. (B) Immunoblotting of cardiomyocytes expressing reduced 14–3–3 (si14–3–3) or normal 14–3–3 (siControl). (C) ChIP–qPCR, showing p-RNAPII S2 at the promoter and internal exon of ANP , β-MHC , GATA-4 , GAPDH and actin in primary neonatal rat cardiomyocytes with siCt or si14–3–3 and stimulated with phenylephrine (PE) for 24 h. (D) ChIP–qPCR assay from mouse heart chromatin, showing p-RNAPII S2 at the ANP , β-MHC , actin and GAPDH exonic region in CaMKII δ -WT or CaMKII δ -KO animals, 21 days after sham operation or TAC. (E) ChIP–qPCR assay, showing p-RNAPII S2 at the ANP , β-MHC , actin and GAPDH promoter regions in CaMKII δ -WT or CaMKII δ -KO animals, 21 days after sham operation or TAC: results are expressed as percentage input over signals with IgG antibody, and represent average ± SD ( n = 3); p values from Student's t -test are indicated

    Techniques Used: Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Binding Assay, Aqueous Normal-phase Chromatography, Mouse Assay, Expressing

    12) Product Images from "Human Myocardial Pericytes: Multipotent Mesodermal Precursors Exhibiting Cardiac Specificity"

    Article Title: Human Myocardial Pericytes: Multipotent Mesodermal Precursors Exhibiting Cardiac Specificity

    Journal: Stem cells (Dayton, Ohio)

    doi: 10.1002/stem.1868

    Cardiomyocyte differentiation of human heart pericytes in vivo
    Figure Legend Snippet: Cardiomyocyte differentiation of human heart pericytes in vivo

    Techniques Used: In Vivo

    Cardiomyocyte differentiation of human heart pericytes in vitro
    Figure Legend Snippet: Cardiomyocyte differentiation of human heart pericytes in vitro

    Techniques Used: In Vitro

    13) Product Images from "Stilbene derivative as a photosensitive compound to control the excitability of neonatal rat cardiomyocytes"

    Article Title: Stilbene derivative as a photosensitive compound to control the excitability of neonatal rat cardiomyocytes

    Journal: Bioscience Reports

    doi: 10.1042/BSR20181849

    Effect of c-TAB on voltage-gated Ca 2+ currents in neonatal rat ventricular cardiomyocytes ( A ) L-type Ca 2+ currents obtained in the absence (control) and presence of 60 μM trans - с-TAB and after UV irradiation. Inset: original current trace in response to a voltage step from −40 mV to 0 mV for 300 ms. Inactivation of INav was achieved by a pre-step from a holding potential HP of −80 mV to −40 mV for 100 ms. Similar results were obtained in more than three additional cells. ( B ) Calcium current ICa, L-type recorded before and after treatment with 60 μM с-TAB and expressed as a percentage of the control ICa, L-type . Each cardiomyocyte was incubated in the presence of с-TAB at room temperature for ∼3 min in a measuring chamber. The data are the means ± SEM, n =5, statistical significance was P
    Figure Legend Snippet: Effect of c-TAB on voltage-gated Ca 2+ currents in neonatal rat ventricular cardiomyocytes ( A ) L-type Ca 2+ currents obtained in the absence (control) and presence of 60 μM trans - с-TAB and after UV irradiation. Inset: original current trace in response to a voltage step from −40 mV to 0 mV for 300 ms. Inactivation of INav was achieved by a pre-step from a holding potential HP of −80 mV to −40 mV for 100 ms. Similar results were obtained in more than three additional cells. ( B ) Calcium current ICa, L-type recorded before and after treatment with 60 μM с-TAB and expressed as a percentage of the control ICa, L-type . Each cardiomyocyte was incubated in the presence of с-TAB at room temperature for ∼3 min in a measuring chamber. The data are the means ± SEM, n =5, statistical significance was P

    Techniques Used: Irradiation, Mass Spectrometry, Incubation

    Effect of trans -c-TAB and UV-light on voltage-gated Nav current in neonatal rat ventricular myocytes ( A ) Effect of c-TAB on ramp currents in neonatal rat ventricular myocytes. Scaled ramp-evoked currents recorded in response to the same ramp protocol (from −120 to +50 mV, 200 ms) in the control and after the addition of 60 μM trans - c-TAB. Three minutes after the application, the current was inhibited by approximately 90% relative to that of the control, and it was not restored after UV irradiation (∼365 nm). Similar results were obtained in more than three additional cells. ( B ) Concentration dependency for trans - c-TAB-induced inhibition of INav in neonatal rat ventricular cardiomyocytes. Mean ± SEM, n =3–4 for each point, statistical significance was P
    Figure Legend Snippet: Effect of trans -c-TAB and UV-light on voltage-gated Nav current in neonatal rat ventricular myocytes ( A ) Effect of c-TAB on ramp currents in neonatal rat ventricular myocytes. Scaled ramp-evoked currents recorded in response to the same ramp protocol (from −120 to +50 mV, 200 ms) in the control and after the addition of 60 μM trans - c-TAB. Three minutes after the application, the current was inhibited by approximately 90% relative to that of the control, and it was not restored after UV irradiation (∼365 nm). Similar results were obtained in more than three additional cells. ( B ) Concentration dependency for trans - c-TAB-induced inhibition of INav in neonatal rat ventricular cardiomyocytes. Mean ± SEM, n =3–4 for each point, statistical significance was P

    Techniques Used: Mass Spectrometry, Irradiation, Concentration Assay, Inhibition

    Effect of c-TAB on voltage-gated K+ currents in neonatal rat ventricular cardiomyocytes ( A ) Whole-cell outward K + currents of the control in response to 500 ms depolarizing voltage steps from −70 mV to +60 mV, after application of 60 μM c-TAB under visible light ( trans -c-TAB) and after ∼365 nm UV irradiation. ( B ) Steady-state K + currents recorded before and after treatment with 60 μM c-TAB and expressed as a percentage of the control IKs. Each cardiomyocyte was incubated in the presence of c-TAB at room temperature for ∼3 min in a measuring chamber. The data are the means ± SEM, n =5, statistical significance was P
    Figure Legend Snippet: Effect of c-TAB on voltage-gated K+ currents in neonatal rat ventricular cardiomyocytes ( A ) Whole-cell outward K + currents of the control in response to 500 ms depolarizing voltage steps from −70 mV to +60 mV, after application of 60 μM c-TAB under visible light ( trans -c-TAB) and after ∼365 nm UV irradiation. ( B ) Steady-state K + currents recorded before and after treatment with 60 μM c-TAB and expressed as a percentage of the control IKs. Each cardiomyocyte was incubated in the presence of c-TAB at room temperature for ∼3 min in a measuring chamber. The data are the means ± SEM, n =5, statistical significance was P

    Techniques Used: Mass Spectrometry, Irradiation, Incubation

    Effect of trans - c-TAB on voltage-gated Na+ currents in neonatal rat ventricular myocytes ( A ) Effect of trans - c-TAB on action potential (AP) formation in rat neonatal myocytes. Superimposed representative traces of APs recorded within the same rat neonatal myocyte in control conditions and during exposure to 60 μM trans- c-TAB. Dynamic of suppression of voltage-gated Na+ current in the presence of trans -c-TAB. Scaled ramp-evoked currents recorded in response to the same ramp protocol (from −120 to +50 mV, 200 ms, HP −80 mV) in the control and after the addition of 60 μM trans- c-TAB. ( C ) Ion currents recorded before and after incubation with 60 μM trans- c-TAB and expressed as percentage. Each cardiomyocyte was incubated in the presence of c-TAB at room temperature for at least 3 min in a measuring chamber. The data represent the means ± SEM, n =6, statistical significance was P
    Figure Legend Snippet: Effect of trans - c-TAB on voltage-gated Na+ currents in neonatal rat ventricular myocytes ( A ) Effect of trans - c-TAB on action potential (AP) formation in rat neonatal myocytes. Superimposed representative traces of APs recorded within the same rat neonatal myocyte in control conditions and during exposure to 60 μM trans- c-TAB. Dynamic of suppression of voltage-gated Na+ current in the presence of trans -c-TAB. Scaled ramp-evoked currents recorded in response to the same ramp protocol (from −120 to +50 mV, 200 ms, HP −80 mV) in the control and after the addition of 60 μM trans- c-TAB. ( C ) Ion currents recorded before and after incubation with 60 μM trans- c-TAB and expressed as percentage. Each cardiomyocyte was incubated in the presence of c-TAB at room temperature for at least 3 min in a measuring chamber. The data represent the means ± SEM, n =6, statistical significance was P

    Techniques Used: Mass Spectrometry, Incubation

    14) Product Images from "Muscle RING finger-1 attenuates IGF-I-dependent cardiomyocyte hypertrophy by inhibiting JNK signaling"

    Article Title: Muscle RING finger-1 attenuates IGF-I-dependent cardiomyocyte hypertrophy by inhibiting JNK signaling

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    doi: 10.1152/ajpendo.00326.2013

    Exaggerated IGF-I-dependent cardiomyocyte growth and Akt-associated gene expression with MuRF1 knockdown requires JNK activity in HL-1 cells. A : MuRF1 was knocked down using AdshMuRF1, with Adshscrambled as control, in HL-1 cells at MOI of 60 for 24 h
    Figure Legend Snippet: Exaggerated IGF-I-dependent cardiomyocyte growth and Akt-associated gene expression with MuRF1 knockdown requires JNK activity in HL-1 cells. A : MuRF1 was knocked down using AdshMuRF1, with Adshscrambled as control, in HL-1 cells at MOI of 60 for 24 h

    Techniques Used: Expressing, Activity Assay

    MuRF1 inhibits cardiomyocyte c-Jun protein levels and phosphorylation in the presence of IGF-I in HL-1 and NRVM cardiomyocytes. A : MuRF1 expression was increased in HL-1 cells using AdMuRF1 and control AdGFP at MOI of 25 for 24 h in serum-free DMEM, followed
    Figure Legend Snippet: MuRF1 inhibits cardiomyocyte c-Jun protein levels and phosphorylation in the presence of IGF-I in HL-1 and NRVM cardiomyocytes. A : MuRF1 expression was increased in HL-1 cells using AdMuRF1 and control AdGFP at MOI of 25 for 24 h in serum-free DMEM, followed

    Techniques Used: Expressing

    IGF-I-dependent expression of genes associated with Akt activity is inhibited by MuRF1 in HL-1 cells. A : AdshMuRF1 and control Adshscrambled were used at MOI of 60 for 24 h in serum-free DMEM to knockdown MuRF1 in HL-1 cardiomyocytes, and transduced cells
    Figure Legend Snippet: IGF-I-dependent expression of genes associated with Akt activity is inhibited by MuRF1 in HL-1 cells. A : AdshMuRF1 and control Adshscrambled were used at MOI of 60 for 24 h in serum-free DMEM to knockdown MuRF1 in HL-1 cardiomyocytes, and transduced cells

    Techniques Used: Expressing, Activity Assay

    Knockdown of muscle RING finger-1 (MuRF1) enhances and increased expression of MuRF1 represses IGF-I-induced cardiomyocyte hypertrophy in HL-1 and neonatal rat ventricular myocytes (NRVM). A : MuRF1 was knocked down using AdshMuRF1 in HL-1 cells, with
    Figure Legend Snippet: Knockdown of muscle RING finger-1 (MuRF1) enhances and increased expression of MuRF1 represses IGF-I-induced cardiomyocyte hypertrophy in HL-1 and neonatal rat ventricular myocytes (NRVM). A : MuRF1 was knocked down using AdshMuRF1 in HL-1 cells, with

    Techniques Used: Expressing

    MuRF1 inhibits protein expression of Akt and GSK-3β in IGF-I-stimulated HL-1 and NRVM cardiomyocytes. A : HL-1 cells were transduced with AdMuRF1 or control AdGFP at MOI of 25 for 24 h in serum-free DMEM to increase MuRF1 expression, followed by
    Figure Legend Snippet: MuRF1 inhibits protein expression of Akt and GSK-3β in IGF-I-stimulated HL-1 and NRVM cardiomyocytes. A : HL-1 cells were transduced with AdMuRF1 or control AdGFP at MOI of 25 for 24 h in serum-free DMEM to increase MuRF1 expression, followed by

    Techniques Used: Expressing, Transduction

    MuRF1 knockdown in HL-1 cardiomyocytes induces total mammalian target of rapamycin (mTOR) protein levels to increase upon IGF-I stimulation. HL-1 cardiomyocytes were transduced with either AdshMuRF1 or control Adshscrambled at MOI of 30 for 48 h in serum-free
    Figure Legend Snippet: MuRF1 knockdown in HL-1 cardiomyocytes induces total mammalian target of rapamycin (mTOR) protein levels to increase upon IGF-I stimulation. HL-1 cardiomyocytes were transduced with either AdshMuRF1 or control Adshscrambled at MOI of 30 for 48 h in serum-free

    Techniques Used: Transduction

    15) Product Images from "ADAP1 limits neonatal cardiomyocyte hypertrophy by reducing integrin cell surface expression"

    Article Title: ADAP1 limits neonatal cardiomyocyte hypertrophy by reducing integrin cell surface expression

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-31784-w

    ADAP1 does not interfere with Mek1ca-induced fetal gene program activation. ( A – I ) Analysis by RT-qPCR of different mRNA expressed in rat neonatal ventricular cardiomyocytes (RNVC) that are representative of the fetal gene program. The RNVC were infected with an β-Gal- (negative control), ADAP1-, or Mek1ca-overexpressing adenovirus, individually or in combination as indicated, and were cultured for 72 h post-infection. The histograms represent mRNA expression levels relative to the β-Gal control and normalized to the Rpl30 reporter gene ( n = 3 independent experiments). * P
    Figure Legend Snippet: ADAP1 does not interfere with Mek1ca-induced fetal gene program activation. ( A – I ) Analysis by RT-qPCR of different mRNA expressed in rat neonatal ventricular cardiomyocytes (RNVC) that are representative of the fetal gene program. The RNVC were infected with an β-Gal- (negative control), ADAP1-, or Mek1ca-overexpressing adenovirus, individually or in combination as indicated, and were cultured for 72 h post-infection. The histograms represent mRNA expression levels relative to the β-Gal control and normalized to the Rpl30 reporter gene ( n = 3 independent experiments). * P

    Techniques Used: Activation Assay, Quantitative RT-PCR, Infection, Negative Control, Cell Culture, Expressing

    ADAP1 restrains the serum-induced increase in cell size of cultured cardiomyocytes. ( A ) Western blot detection of adenovirus-mediated 3xFLAG-hADAP1 overexpression (MOI of 50) in rat neonatal ventricular cardiomyocytes (RNVC) cultured for 72 h post-infection. ( B ) RNVC were infected with either β-Gal- (negative control) or ADAP1-overexpressing adenovirus and were cultured for 72 h in the absence (0%) or presence (10%) of serum. Representative images of α-Actinin-immunostained RNVC (left) and corresponding segmented images were acquired using the Operetta High-Content Imaging System (Perkin Elmer). The scale bar represents 50 µm. ( C ) The histogram represents the cell surface areas of RNVC overexpressing either β-Gal or ADAP1 and cultured for 72 h with increasing concentrations of serum ( n = 3 independent experiments). * P
    Figure Legend Snippet: ADAP1 restrains the serum-induced increase in cell size of cultured cardiomyocytes. ( A ) Western blot detection of adenovirus-mediated 3xFLAG-hADAP1 overexpression (MOI of 50) in rat neonatal ventricular cardiomyocytes (RNVC) cultured for 72 h post-infection. ( B ) RNVC were infected with either β-Gal- (negative control) or ADAP1-overexpressing adenovirus and were cultured for 72 h in the absence (0%) or presence (10%) of serum. Representative images of α-Actinin-immunostained RNVC (left) and corresponding segmented images were acquired using the Operetta High-Content Imaging System (Perkin Elmer). The scale bar represents 50 µm. ( C ) The histogram represents the cell surface areas of RNVC overexpressing either β-Gal or ADAP1 and cultured for 72 h with increasing concentrations of serum ( n = 3 independent experiments). * P

    Techniques Used: Cell Culture, Western Blot, Over Expression, Infection, Negative Control, Imaging

    ADAP1 blocks phenylephrine- and Mek1ca-induced hypertrophy. Cell surface area measurements of rat neonatal ventricular cardiomyocytes (RNVC) overexpressing ADAP1 in the absence or presence of 50 µM phenylephrine ( A ) or Mek1ca-overexpressing adenovirus ( B ) compared with an adβ-Gal-infected control. Quantification of at least 3 independent experiments expressed as means ± SD. * P
    Figure Legend Snippet: ADAP1 blocks phenylephrine- and Mek1ca-induced hypertrophy. Cell surface area measurements of rat neonatal ventricular cardiomyocytes (RNVC) overexpressing ADAP1 in the absence or presence of 50 µM phenylephrine ( A ) or Mek1ca-overexpressing adenovirus ( B ) compared with an adβ-Gal-infected control. Quantification of at least 3 independent experiments expressed as means ± SD. * P

    Techniques Used: Infection

    ADAP1 relocalizes cytoskeletal α-Actinin. ( A ) Representative confocal images (Olympus FluoView FV1000 microscope) of α-Actinin-immunostained rat neonatal ventricular cardiomyocytes (RNVC) infected with a β-Gal- (negative control), ADAP1-, or Mek1ca-overexpressing adenovirus, individually or in combination as indicated, and cultured for 72 h post-infection. Arrows point to α-Actinin dense puncta. The scale bar represents 12 µm. ( B ) Number of α-Actinin puncta per cell measured with the Operetta High-Content Imaging System (Perkin Elmer) using the same experimental conditions as in A ( n = 4 independent experiments). **** P
    Figure Legend Snippet: ADAP1 relocalizes cytoskeletal α-Actinin. ( A ) Representative confocal images (Olympus FluoView FV1000 microscope) of α-Actinin-immunostained rat neonatal ventricular cardiomyocytes (RNVC) infected with a β-Gal- (negative control), ADAP1-, or Mek1ca-overexpressing adenovirus, individually or in combination as indicated, and cultured for 72 h post-infection. Arrows point to α-Actinin dense puncta. The scale bar represents 12 µm. ( B ) Number of α-Actinin puncta per cell measured with the Operetta High-Content Imaging System (Perkin Elmer) using the same experimental conditions as in A ( n = 4 independent experiments). **** P

    Techniques Used: Microscopy, Infection, Negative Control, Cell Culture, Imaging

    Adap1 expression in cardiac cells. ( A ) Relative levels of Adap1 mRNA expression in the rat brain (adult) and heart (adult and 2-day-old neonate) were measured by RT-qPCR and were normalized to the Rpl30 reporter gene ( n = 4 independent tissues). ( B ) Representative Western blots of Adap1 and Gapdh (loading control) detected from whole brain and heart extracts. ( C ) The histogram represents the relative expression level of Adap1 normalized to Gapdh in the respective tissues ( n = 4 independent tissues). ( D ) Representative Western blot of Adap1, α-Actinin (cardiomyocyte specific marker), and Gapdh (loading control) detected in the protein lysates of enriched rat neonatal ventricular cardiomyocytes (RNVC) and non-cardiomyocytes (Non-CM). ( E ) The histogram represents the relative expression level of Adap1 normalized to Gapdh in the respective cell lysates ( n .
    Figure Legend Snippet: Adap1 expression in cardiac cells. ( A ) Relative levels of Adap1 mRNA expression in the rat brain (adult) and heart (adult and 2-day-old neonate) were measured by RT-qPCR and were normalized to the Rpl30 reporter gene ( n = 4 independent tissues). ( B ) Representative Western blots of Adap1 and Gapdh (loading control) detected from whole brain and heart extracts. ( C ) The histogram represents the relative expression level of Adap1 normalized to Gapdh in the respective tissues ( n = 4 independent tissues). ( D ) Representative Western blot of Adap1, α-Actinin (cardiomyocyte specific marker), and Gapdh (loading control) detected in the protein lysates of enriched rat neonatal ventricular cardiomyocytes (RNVC) and non-cardiomyocytes (Non-CM). ( E ) The histogram represents the relative expression level of Adap1 normalized to Gapdh in the respective cell lysates ( n .

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Marker

    16) Product Images from "Docosahexaenoic acid inhibits protein kinase C translocation/activation and cardiac hypertrophy in rat cardiomyocytes"

    Article Title: Docosahexaenoic acid inhibits protein kinase C translocation/activation and cardiac hypertrophy in rat cardiomyocytes

    Journal: Journal of molecular and genetic medicine : an international journal of biomedical research

    doi:

    Effect of PE and DHA on α-actinin and ANF expression. Cardiomyocytes were treated with PE and DHA as described in the legend of Figure 1 . Solid arrows indicate the expression of sarcomeric α-actinin in z-bands (green fluorescence) whereas dotted arrows indicate expression of ANF (red fluorescence). Results are a typical representation of five experiments.
    Figure Legend Snippet: Effect of PE and DHA on α-actinin and ANF expression. Cardiomyocytes were treated with PE and DHA as described in the legend of Figure 1 . Solid arrows indicate the expression of sarcomeric α-actinin in z-bands (green fluorescence) whereas dotted arrows indicate expression of ANF (red fluorescence). Results are a typical representation of five experiments.

    Techniques Used: Expressing, Fluorescence

    Effect of PE and DHA on PKCα translocation in cardiomyocytes. Cardiomyocytes were treated with PE and DHA as described in the legend of Figure 1 . Proteins were detected using anti-PKCα specific antibody and visualized with Alexa 546- (red fluorescence) labelled anti-mouse as described in the Material and Methods section. Cells were examined under a fluorescence microscope and images were captured using a MagnaFire digital camera (Optronics) for analysis. Results are a typical representation of three experiments.
    Figure Legend Snippet: Effect of PE and DHA on PKCα translocation in cardiomyocytes. Cardiomyocytes were treated with PE and DHA as described in the legend of Figure 1 . Proteins were detected using anti-PKCα specific antibody and visualized with Alexa 546- (red fluorescence) labelled anti-mouse as described in the Material and Methods section. Cells were examined under a fluorescence microscope and images were captured using a MagnaFire digital camera (Optronics) for analysis. Results are a typical representation of three experiments.

    Techniques Used: Translocation Assay, Fluorescence, Microscopy

    Effect of PE and DHA on cell surface area. Cardiomyocytes were treated with either ethanol (control), serum free media or DHA (5 μM in serum-free conditions) for 24 hr and then incubated with/without PE (100 μM) for 48 hr. Data are expressed as mean ± SEM for examination of 10 cells in each group from three separate experiments and were analyzed by ANOVA and Tukey's multiple comparison tests. Significant differences within groups are reported. ***P
    Figure Legend Snippet: Effect of PE and DHA on cell surface area. Cardiomyocytes were treated with either ethanol (control), serum free media or DHA (5 μM in serum-free conditions) for 24 hr and then incubated with/without PE (100 μM) for 48 hr. Data are expressed as mean ± SEM for examination of 10 cells in each group from three separate experiments and were analyzed by ANOVA and Tukey's multiple comparison tests. Significant differences within groups are reported. ***P

    Techniques Used: Incubation

    Effect of PE and DHA on PKC activity. Cardiomyocytes were treated with PE and DHA as described in the legend of Figure 1 . The total PKC activity in membrane fractions of cardiomyocytes was assayed as described in the Material and Methods section. Results are expressed as the mean±SE for three experiments and analyzed by ANOVA and Tukey's multiple comparison tests. Significant differences within groups are reported. ***P
    Figure Legend Snippet: Effect of PE and DHA on PKC activity. Cardiomyocytes were treated with PE and DHA as described in the legend of Figure 1 . The total PKC activity in membrane fractions of cardiomyocytes was assayed as described in the Material and Methods section. Results are expressed as the mean±SE for three experiments and analyzed by ANOVA and Tukey's multiple comparison tests. Significant differences within groups are reported. ***P

    Techniques Used: Activity Assay

    Effect of PE and DHA on PKCα distribution in membranes. Cardiomyocytes were treated with PE and DHA as described in the legend of Figure 1 . Membranes were isolated and separated on 8% SDS-PAGE as described in the Material and Methods section. PKCα was detected by Western blotting using anti-PKCα antibodies and the relative distribution of PKCα was determined by densitometric analysis. Results are a typical representation of three experiments.
    Figure Legend Snippet: Effect of PE and DHA on PKCα distribution in membranes. Cardiomyocytes were treated with PE and DHA as described in the legend of Figure 1 . Membranes were isolated and separated on 8% SDS-PAGE as described in the Material and Methods section. PKCα was detected by Western blotting using anti-PKCα antibodies and the relative distribution of PKCα was determined by densitometric analysis. Results are a typical representation of three experiments.

    Techniques Used: Isolation, SDS Page, Western Blot

    17) Product Images from "Construction of Cardiac Tissue Rings Using a Magnetic Tissue Fabrication Technique"

    Article Title: Construction of Cardiac Tissue Rings Using a Magnetic Tissue Fabrication Technique

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms11082910

    A procedure for fabrication of a cardiac tissue ring by combining Mag-TE and ECM-based techniques. A mixture of diluted ECM precursor and MCL-labeled cardiomyocytes was cast into a well of a 24-well ultra-low attachment plate containing a polycarbonate cylinder fixed in the center of each well. Immediately thereafter, a magnet was placed underneath the wells to attract the MCL-labeled cardiomyocytes to the culture bottom, enabling the removal of excess amounts of ECM precursor from the upper side of the formed cell layer. The remaining ECM within the cell layer was then hardened, and the medium was added. During the culture, the cell layer gradually shrank towards the cylinder, resulting in the formation of ring-shaped cardiac tissue.
    Figure Legend Snippet: A procedure for fabrication of a cardiac tissue ring by combining Mag-TE and ECM-based techniques. A mixture of diluted ECM precursor and MCL-labeled cardiomyocytes was cast into a well of a 24-well ultra-low attachment plate containing a polycarbonate cylinder fixed in the center of each well. Immediately thereafter, a magnet was placed underneath the wells to attract the MCL-labeled cardiomyocytes to the culture bottom, enabling the removal of excess amounts of ECM precursor from the upper side of the formed cell layer. The remaining ECM within the cell layer was then hardened, and the medium was added. During the culture, the cell layer gradually shrank towards the cylinder, resulting in the formation of ring-shaped cardiac tissue.

    Techniques Used: Labeling

    MCL-labeling of cardiomyocytes. ( a ) Measurement of MCL amount captured by the cells. Isolated cardiomyocytes were suspended in the medium containing various MCL concentrations (0, 25, 50, 100 and 200 pg/cell), and incubated for 1 h at 4 °C. Subsequently, the magnetite amount captured by the cells was measured using the potassium thiocyanate method. ( b ) The effect of MCL-labeling on cell viability. Cardiomyocytes with or without MCL-labeling were seeded into wells of a 96-well plate. After 4-day culture, cell viability was measured using the WST-8 assay. Data are expressed as mean ± SD (n = 3). * P
    Figure Legend Snippet: MCL-labeling of cardiomyocytes. ( a ) Measurement of MCL amount captured by the cells. Isolated cardiomyocytes were suspended in the medium containing various MCL concentrations (0, 25, 50, 100 and 200 pg/cell), and incubated for 1 h at 4 °C. Subsequently, the magnetite amount captured by the cells was measured using the potassium thiocyanate method. ( b ) The effect of MCL-labeling on cell viability. Cardiomyocytes with or without MCL-labeling were seeded into wells of a 96-well plate. After 4-day culture, cell viability was measured using the WST-8 assay. Data are expressed as mean ± SD (n = 3). * P

    Techniques Used: Labeling, Isolation, Incubation

    18) Product Images from "Early cardiac hypertrophy in mice with impaired calmodulin regulation of cardiac muscle Ca2+ release channel"

    Article Title: Early cardiac hypertrophy in mice with impaired calmodulin regulation of cardiac muscle Ca2+ release channel

    Journal:

    doi: 10.1172/JCI29515

    Ca 2+ imaging of cardiomyocytes from WT and mutant hearts.
    Figure Legend Snippet: Ca 2+ imaging of cardiomyocytes from WT and mutant hearts.

    Techniques Used: Imaging, Mutagenesis

    19) Product Images from "Control of histone H3 phosphorylation by CaMKIIδ in response to haemodynamic cardiac stress"

    Article Title: Control of histone H3 phosphorylation by CaMKIIδ in response to haemodynamic cardiac stress

    Journal: The Journal of Pathology

    doi: 10.1002/path.4489

    14–3–3 recruitment to chromatin is important for transcriptional elongation of GATA-4 and of fetal cardiac genes. (A) ChIP–qPCR performed from mouse heart tissues showing 14–3–3 binding at the ANP and β -MHC promoters in CaMKII δ -WT or CaMKII δ -KO mice 21 days after sham operation or TAC. qPCR reactions were also performed at GAPDH and α-actin promoters that are not responsive to pressure overload hypertrophy: results are expressed as percentage input over signals with IgG antibody and represent average ± SD ( n = 3); p values from Student's t test are indicated. (B) Immunoblotting of cardiomyocytes expressing reduced 14–3–3 (si14–3–3) or normal 14–3–3 (siControl). (C) ChIP–qPCR, showing p-RNAPII S2 at the promoter and internal exon of ANP , β-MHC , GATA-4 , GAPDH and actin in primary neonatal rat cardiomyocytes with siCt or si14–3–3 and stimulated with phenylephrine (PE) for 24 h. (D) ChIP–qPCR assay from mouse heart chromatin, showing p-RNAPII S2 at the ANP , β-MHC , actin and GAPDH exonic region in CaMKII δ -WT or CaMKII δ -KO animals, 21 days after sham operation or TAC. (E) ChIP–qPCR assay, showing p-RNAPII S2 at the ANP , β-MHC , actin and GAPDH promoter regions in CaMKII δ -WT or CaMKII δ -KO animals, 21 days after sham operation or TAC: results are expressed as percentage input over signals with IgG antibody, and represent average ± SD ( n = 3); p values from Student's t -test are indicated
    Figure Legend Snippet: 14–3–3 recruitment to chromatin is important for transcriptional elongation of GATA-4 and of fetal cardiac genes. (A) ChIP–qPCR performed from mouse heart tissues showing 14–3–3 binding at the ANP and β -MHC promoters in CaMKII δ -WT or CaMKII δ -KO mice 21 days after sham operation or TAC. qPCR reactions were also performed at GAPDH and α-actin promoters that are not responsive to pressure overload hypertrophy: results are expressed as percentage input over signals with IgG antibody and represent average ± SD ( n = 3); p values from Student's t test are indicated. (B) Immunoblotting of cardiomyocytes expressing reduced 14–3–3 (si14–3–3) or normal 14–3–3 (siControl). (C) ChIP–qPCR, showing p-RNAPII S2 at the promoter and internal exon of ANP , β-MHC , GATA-4 , GAPDH and actin in primary neonatal rat cardiomyocytes with siCt or si14–3–3 and stimulated with phenylephrine (PE) for 24 h. (D) ChIP–qPCR assay from mouse heart chromatin, showing p-RNAPII S2 at the ANP , β-MHC , actin and GAPDH exonic region in CaMKII δ -WT or CaMKII δ -KO animals, 21 days after sham operation or TAC. (E) ChIP–qPCR assay, showing p-RNAPII S2 at the ANP , β-MHC , actin and GAPDH promoter regions in CaMKII δ -WT or CaMKII δ -KO animals, 21 days after sham operation or TAC: results are expressed as percentage input over signals with IgG antibody, and represent average ± SD ( n = 3); p values from Student's t -test are indicated

    Techniques Used: Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Binding Assay, Aqueous Normal-phase Chromatography, Mouse Assay, Expressing

    20) Product Images from "Spatiotemporal Stability of Neonatal Rat Cardiomyocyte Monolayers Spontaneous Activity Is Dependent on the Culture Substrate"

    Article Title: Spatiotemporal Stability of Neonatal Rat Cardiomyocyte Monolayers Spontaneous Activity Is Dependent on the Culture Substrate

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0127977

    Role of the proteins expression on spontaneous activity. A. Cx43 mRNA expression was 6.24±0.89, 4.55±0.27 and 5.70±0.12. According to our results, Cx43 mRNA expression has a tendency to be lower on PDMS substrates compared to glass, however, significant differences was detected between PDMS 1:20 and PDMS 1:40. B-D. mRNA expression of targeted proteins playing a role on automaticity through the voltage clock could explain the small changes observed in rhythm. Kir2.1 (B) mRNA expression tends to be lowered when cardiomyocytes are cultivated on PDMS compared to glass. There is a tendency towards increased expression of CaV3.1 (C) and HCN2 (D) on PDMS compared to glass (p
    Figure Legend Snippet: Role of the proteins expression on spontaneous activity. A. Cx43 mRNA expression was 6.24±0.89, 4.55±0.27 and 5.70±0.12. According to our results, Cx43 mRNA expression has a tendency to be lower on PDMS substrates compared to glass, however, significant differences was detected between PDMS 1:20 and PDMS 1:40. B-D. mRNA expression of targeted proteins playing a role on automaticity through the voltage clock could explain the small changes observed in rhythm. Kir2.1 (B) mRNA expression tends to be lowered when cardiomyocytes are cultivated on PDMS compared to glass. There is a tendency towards increased expression of CaV3.1 (C) and HCN2 (D) on PDMS compared to glass (p

    Techniques Used: Expressing, Activity Assay

    Increased number of initiation sites on PDMS substrate. A. The mean number of activation sites for cardiomyocyte monolayers cultivated on glass, PDMS 1:20, and PDMS 1:40 were 1.3±0.3, 2.8±0.4, and 2.8±0.5 sites, respectively. The mean number of activation sites is significantly higher on PDMS compared to glass (p
    Figure Legend Snippet: Increased number of initiation sites on PDMS substrate. A. The mean number of activation sites for cardiomyocyte monolayers cultivated on glass, PDMS 1:20, and PDMS 1:40 were 1.3±0.3, 2.8±0.4, and 2.8±0.5 sites, respectively. The mean number of activation sites is significantly higher on PDMS compared to glass (p

    Techniques Used: Activation Assay

    ISO decreases σ period for cardiomyocytes cultivated on PDMS 1:20 and tends to stabilize the rate of contraction. Examples of cardiomyocyte activity stabilization by the addition of isoproterenol. i) A trace of the contractile activity is shown with ii) activation maps of the first beat for each different activation site. A . Conditions before the addition of isoproterenol (Pre-ISO) on PDMS 1:20. B . One minute after adding ISO (100 nM) on PDMS 1:20 substrates. Pharmacological sympathetic stimulation appears to decrease the number of activation sites (from 4 sites pre-ISO to 2 sites after ISO).
    Figure Legend Snippet: ISO decreases σ period for cardiomyocytes cultivated on PDMS 1:20 and tends to stabilize the rate of contraction. Examples of cardiomyocyte activity stabilization by the addition of isoproterenol. i) A trace of the contractile activity is shown with ii) activation maps of the first beat for each different activation site. A . Conditions before the addition of isoproterenol (Pre-ISO) on PDMS 1:20. B . One minute after adding ISO (100 nM) on PDMS 1:20 substrates. Pharmacological sympathetic stimulation appears to decrease the number of activation sites (from 4 sites pre-ISO to 2 sites after ISO).

    Techniques Used: Activity Assay, Activation Assay

    21) Product Images from "Cardiomyocyte Overexpression of FABP4 Aggravates Pressure Overload-Induced Heart Hypertrophy"

    Article Title: Cardiomyocyte Overexpression of FABP4 Aggravates Pressure Overload-Induced Heart Hypertrophy

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0157372

    Generation of cardiomyocyte-specific FABP4 transgenic mice. (A) Schematics of mouse α-MHC-driven FABP4 transgenic construct, HGH-PA: human growth hormone polyadenylation signal. (B) The expression of FABP4 in isolated adult cardiomyocytes from wild type and FABP4-TG mice. Relative expression levels of FABP4 protein normalized by GAPDH are expressed as mean±SD (n = 6, **p
    Figure Legend Snippet: Generation of cardiomyocyte-specific FABP4 transgenic mice. (A) Schematics of mouse α-MHC-driven FABP4 transgenic construct, HGH-PA: human growth hormone polyadenylation signal. (B) The expression of FABP4 in isolated adult cardiomyocytes from wild type and FABP4-TG mice. Relative expression levels of FABP4 protein normalized by GAPDH are expressed as mean±SD (n = 6, **p

    Techniques Used: Transgenic Assay, Mouse Assay, Construct, Expressing, Isolation

    22) Product Images from "Actin-dependent mitochondrial internalization in cardiomyocytes: evidence for rescue of mitochondrial function"

    Article Title: Actin-dependent mitochondrial internalization in cardiomyocytes: evidence for rescue of mitochondrial function

    Journal: Biology Open

    doi: 10.1242/bio.201511478

    Mitochondrial internalization in cardiomyocytes. (A) Representative florescent micrographs of isolated rat liver mitochondria labeled with pHrodo co-incubated with cardiomyocytes for 4 hour (middle panel) or 24 hours (right panel). Control cardiomyocytes (no mitochondria) are shown in the left panel. In all images the blue stain is DAPI; red, pHrodo labeled mitochondria, green is 488 phalloidin (f-actin). Scale bars are 25 µm. Results show internalization of transplanted mitochondria at 4 and 24 hours. (B) ATP content in cardiomyocytes: ATP content (nmol/10 3 cells) in control, no mitochondria (white bars) and cardiomyocytes co-incubated with mitochondria (black bars). (C) Mitochondrial uptake at 1, 4 and 24 hour co-incubation. The mean±sd for n = 5 experiments is shown. Results indicate that ATP content is significantly increased (p
    Figure Legend Snippet: Mitochondrial internalization in cardiomyocytes. (A) Representative florescent micrographs of isolated rat liver mitochondria labeled with pHrodo co-incubated with cardiomyocytes for 4 hour (middle panel) or 24 hours (right panel). Control cardiomyocytes (no mitochondria) are shown in the left panel. In all images the blue stain is DAPI; red, pHrodo labeled mitochondria, green is 488 phalloidin (f-actin). Scale bars are 25 µm. Results show internalization of transplanted mitochondria at 4 and 24 hours. (B) ATP content in cardiomyocytes: ATP content (nmol/10 3 cells) in control, no mitochondria (white bars) and cardiomyocytes co-incubated with mitochondria (black bars). (C) Mitochondrial uptake at 1, 4 and 24 hour co-incubation. The mean±sd for n = 5 experiments is shown. Results indicate that ATP content is significantly increased (p

    Techniques Used: Isolation, Labeling, Incubation, Staining

    Inhibition of mitochondrial internalization. (A) Representative florescent micrographs of 2-day cardiomyocytes co-incubated with 1×10 7 rat liver mitochondria labeled with pHrodo red following 4 hours co-incubation. Actin-dependent endocytosis and phagocytosis was inhibited with cytochalasin D (CytoD), caveola-dependent-clathrin dependent endocytosis was inhibited with methyl-β–cyclodextrin (MβCD), tunneling nano tube formation was inhibited with nocodazole (Noco) and macro pinocytosis was blocked with 5-(N-Ethyl-N-isopropyl)amiloride (EIPA) at 10 µM, 50 µM or 100 µM . Control cardiomyocytes were co-incubated with mitochondria without inhibitor (no inhibitor), As separate control, cardiomyocytes without mitochondria (no mitochondria) is also shown. Internalized mitochondria are red (pHrodo labeled mitochondria). Nuclei are shown in blue (DAPI). Scale bars are 25 µm. (B) Internalization of mitochondria per nucleus as determined by florescent microscopy following 4 hours of co-incubation. (C) ATP content (nmol/10 3 cells). Results are shown as the mean and standard deviation for n = 5 in each group. Statistical differences at p
    Figure Legend Snippet: Inhibition of mitochondrial internalization. (A) Representative florescent micrographs of 2-day cardiomyocytes co-incubated with 1×10 7 rat liver mitochondria labeled with pHrodo red following 4 hours co-incubation. Actin-dependent endocytosis and phagocytosis was inhibited with cytochalasin D (CytoD), caveola-dependent-clathrin dependent endocytosis was inhibited with methyl-β–cyclodextrin (MβCD), tunneling nano tube formation was inhibited with nocodazole (Noco) and macro pinocytosis was blocked with 5-(N-Ethyl-N-isopropyl)amiloride (EIPA) at 10 µM, 50 µM or 100 µM . Control cardiomyocytes were co-incubated with mitochondria without inhibitor (no inhibitor), As separate control, cardiomyocytes without mitochondria (no mitochondria) is also shown. Internalized mitochondria are red (pHrodo labeled mitochondria). Nuclei are shown in blue (DAPI). Scale bars are 25 µm. (B) Internalization of mitochondria per nucleus as determined by florescent microscopy following 4 hours of co-incubation. (C) ATP content (nmol/10 3 cells). Results are shown as the mean and standard deviation for n = 5 in each group. Statistical differences at p

    Techniques Used: Inhibition, Incubation, Labeling, Microscopy, Standard Deviation

    23) Product Images from "Cardiac Niche Influences the Direct Reprogramming of Canine Fibroblasts into Cardiomyocyte-Like Cells"

    Article Title: Cardiac Niche Influences the Direct Reprogramming of Canine Fibroblasts into Cardiomyocyte-Like Cells

    Journal: Stem Cells International

    doi: 10.1155/2016/4969430

    Viral transduction and exogenous gene expressions. (a) Schematic representation of the lentiviral vectors carrying the human sequence of the transcription factor genes (GATA4-MEF2C-TBX5-HAND2), the microdystrophin gene, and the GFP reporter gene. (b) The GFP reporter gene under the control of the α -MyHC promoter was already expressed 48 hours after transduction. (c) Fibroblasts underwent morphology changes 7 days after transduction and become flattened and binucleated cells expressing GFP. (d) Immunofluorescence analysis on GHMT CF alone (top left panel) and in coculture with neonatal rat cardiomyocytes (bottom left panel), with antibodies against GFP (green) and myosin heavy chain (red), shows double positive cells and single positive cells for GFP (right panel). (e) Cell growth curve after transduction shows a decrease of cell proliferation ( n = 5/cell type). (f) Fold induction of microdystrophin ( μ dys) mRNA expression in transduced cells compared to the control; n = 5 ∗ p
    Figure Legend Snippet: Viral transduction and exogenous gene expressions. (a) Schematic representation of the lentiviral vectors carrying the human sequence of the transcription factor genes (GATA4-MEF2C-TBX5-HAND2), the microdystrophin gene, and the GFP reporter gene. (b) The GFP reporter gene under the control of the α -MyHC promoter was already expressed 48 hours after transduction. (c) Fibroblasts underwent morphology changes 7 days after transduction and become flattened and binucleated cells expressing GFP. (d) Immunofluorescence analysis on GHMT CF alone (top left panel) and in coculture with neonatal rat cardiomyocytes (bottom left panel), with antibodies against GFP (green) and myosin heavy chain (red), shows double positive cells and single positive cells for GFP (right panel). (e) Cell growth curve after transduction shows a decrease of cell proliferation ( n = 5/cell type). (f) Fold induction of microdystrophin ( μ dys) mRNA expression in transduced cells compared to the control; n = 5 ∗ p

    Techniques Used: Transduction, Sequencing, Expressing, Immunofluorescence

    24) Product Images from "Tauroursodeoxycholic acid (TUDCA) attenuates pressure overload-induced cardiac remodeling by reducing endoplasmic reticulum stress"

    Article Title: Tauroursodeoxycholic acid (TUDCA) attenuates pressure overload-induced cardiac remodeling by reducing endoplasmic reticulum stress

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0176071

    Tauroursodeoxycholic acid (TUDCA) administration reduced the number of apoptotic cells in transverse aortic constriction (TAC)-induced hypertrophic hearts. (A) Protein expression of the pro-apoptotic genes CHOP and cleaved caspase 3 after 4 weeks of TUDCA administration. (B, C) Relative expression of the pro-apoptotic genes CHOP and cleaved caspase 3 ( n = 3). (D) A TUNEL assay was performed using paraffin-embedded heart tissues (scale bar = 50 μm). (E) Quantitative representation of the percentage of TUNEL-positive cardiomyocytes. Only nuclei that were purplish red were scored. The percentage of TUNEL-positive cells was calculated as the ratio of TUNEL-positive to DAPI-stained nuclei in the section. Three to five fields were selected randomly for each heart section, and quantitated by ImageJ ( n = 3–6). All data are shown as mean ± SE (* P
    Figure Legend Snippet: Tauroursodeoxycholic acid (TUDCA) administration reduced the number of apoptotic cells in transverse aortic constriction (TAC)-induced hypertrophic hearts. (A) Protein expression of the pro-apoptotic genes CHOP and cleaved caspase 3 after 4 weeks of TUDCA administration. (B, C) Relative expression of the pro-apoptotic genes CHOP and cleaved caspase 3 ( n = 3). (D) A TUNEL assay was performed using paraffin-embedded heart tissues (scale bar = 50 μm). (E) Quantitative representation of the percentage of TUNEL-positive cardiomyocytes. Only nuclei that were purplish red were scored. The percentage of TUNEL-positive cells was calculated as the ratio of TUNEL-positive to DAPI-stained nuclei in the section. Three to five fields were selected randomly for each heart section, and quantitated by ImageJ ( n = 3–6). All data are shown as mean ± SE (* P

    Techniques Used: Expressing, TUNEL Assay, Staining

    25) Product Images from "ADAP1 limits neonatal cardiomyocyte hypertrophy by reducing integrin cell surface expression"

    Article Title: ADAP1 limits neonatal cardiomyocyte hypertrophy by reducing integrin cell surface expression

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-31784-w

    ADAP1 does not interfere with Mek1ca-induced fetal gene program activation. ( A – I ) Analysis by RT-qPCR of different mRNA expressed in rat neonatal ventricular cardiomyocytes (RNVC) that are representative of the fetal gene program. The RNVC were infected with an β-Gal- (negative control), ADAP1-, or Mek1ca-overexpressing adenovirus, individually or in combination as indicated, and were cultured for 72 h post-infection. The histograms represent mRNA expression levels relative to the β-Gal control and normalized to the Rpl30 reporter gene ( n = 3 independent experiments). * P
    Figure Legend Snippet: ADAP1 does not interfere with Mek1ca-induced fetal gene program activation. ( A – I ) Analysis by RT-qPCR of different mRNA expressed in rat neonatal ventricular cardiomyocytes (RNVC) that are representative of the fetal gene program. The RNVC were infected with an β-Gal- (negative control), ADAP1-, or Mek1ca-overexpressing adenovirus, individually or in combination as indicated, and were cultured for 72 h post-infection. The histograms represent mRNA expression levels relative to the β-Gal control and normalized to the Rpl30 reporter gene ( n = 3 independent experiments). * P

    Techniques Used: Activation Assay, Quantitative RT-PCR, Infection, Negative Control, Cell Culture, Expressing

    ADAP1 restrains the serum-induced increase in cell size of cultured cardiomyocytes. ( A ) Western blot detection of adenovirus-mediated 3xFLAG-hADAP1 overexpression (MOI of 50) in rat neonatal ventricular cardiomyocytes (RNVC) cultured for 72 h post-infection. ( B ) RNVC were infected with either β-Gal- (negative control) or ADAP1-overexpressing adenovirus and were cultured for 72 h in the absence (0%) or presence (10%) of serum. Representative images of α-Actinin-immunostained RNVC (left) and corresponding segmented images were acquired using the Operetta High-Content Imaging System (Perkin Elmer). The scale bar represents 50 µm. ( C ) The histogram represents the cell surface areas of RNVC overexpressing either β-Gal or ADAP1 and cultured for 72 h with increasing concentrations of serum ( n = 3 independent experiments). * P
    Figure Legend Snippet: ADAP1 restrains the serum-induced increase in cell size of cultured cardiomyocytes. ( A ) Western blot detection of adenovirus-mediated 3xFLAG-hADAP1 overexpression (MOI of 50) in rat neonatal ventricular cardiomyocytes (RNVC) cultured for 72 h post-infection. ( B ) RNVC were infected with either β-Gal- (negative control) or ADAP1-overexpressing adenovirus and were cultured for 72 h in the absence (0%) or presence (10%) of serum. Representative images of α-Actinin-immunostained RNVC (left) and corresponding segmented images were acquired using the Operetta High-Content Imaging System (Perkin Elmer). The scale bar represents 50 µm. ( C ) The histogram represents the cell surface areas of RNVC overexpressing either β-Gal or ADAP1 and cultured for 72 h with increasing concentrations of serum ( n = 3 independent experiments). * P

    Techniques Used: Cell Culture, Western Blot, Over Expression, Infection, Negative Control, Imaging

    ADAP1 blocks phenylephrine- and Mek1ca-induced hypertrophy. Cell surface area measurements of rat neonatal ventricular cardiomyocytes (RNVC) overexpressing ADAP1 in the absence or presence of 50 µM phenylephrine ( A ) or Mek1ca-overexpressing adenovirus ( B ) compared with an adβ-Gal-infected control. Quantification of at least 3 independent experiments expressed as means ± SD. * P
    Figure Legend Snippet: ADAP1 blocks phenylephrine- and Mek1ca-induced hypertrophy. Cell surface area measurements of rat neonatal ventricular cardiomyocytes (RNVC) overexpressing ADAP1 in the absence or presence of 50 µM phenylephrine ( A ) or Mek1ca-overexpressing adenovirus ( B ) compared with an adβ-Gal-infected control. Quantification of at least 3 independent experiments expressed as means ± SD. * P

    Techniques Used: Infection

    ADAP1 relocalizes cytoskeletal α-Actinin. ( A ) Representative confocal images (Olympus FluoView FV1000 microscope) of α-Actinin-immunostained rat neonatal ventricular cardiomyocytes (RNVC) infected with a β-Gal- (negative control), ADAP1-, or Mek1ca-overexpressing adenovirus, individually or in combination as indicated, and cultured for 72 h post-infection. Arrows point to α-Actinin dense puncta. The scale bar represents 12 µm. ( B ) Number of α-Actinin puncta per cell measured with the Operetta High-Content Imaging System (Perkin Elmer) using the same experimental conditions as in A ( n = 4 independent experiments). **** P
    Figure Legend Snippet: ADAP1 relocalizes cytoskeletal α-Actinin. ( A ) Representative confocal images (Olympus FluoView FV1000 microscope) of α-Actinin-immunostained rat neonatal ventricular cardiomyocytes (RNVC) infected with a β-Gal- (negative control), ADAP1-, or Mek1ca-overexpressing adenovirus, individually or in combination as indicated, and cultured for 72 h post-infection. Arrows point to α-Actinin dense puncta. The scale bar represents 12 µm. ( B ) Number of α-Actinin puncta per cell measured with the Operetta High-Content Imaging System (Perkin Elmer) using the same experimental conditions as in A ( n = 4 independent experiments). **** P

    Techniques Used: Microscopy, Infection, Negative Control, Cell Culture, Imaging

    Adap1 expression in cardiac cells. ( A ) Relative levels of Adap1 mRNA expression in the rat brain (adult) and heart (adult and 2-day-old neonate) were measured by RT-qPCR and were normalized to the Rpl30 reporter gene ( n = 4 independent tissues). ( B ) Representative Western blots of Adap1 and Gapdh (loading control) detected from whole brain and heart extracts. ( C ) The histogram represents the relative expression level of Adap1 normalized to Gapdh in the respective tissues ( n = 4 independent tissues). ( D ) Representative Western blot of Adap1, α-Actinin (cardiomyocyte specific marker), and Gapdh (loading control) detected in the protein lysates of enriched rat neonatal ventricular cardiomyocytes (RNVC) and non-cardiomyocytes (Non-CM). ( E ) The histogram represents the relative expression level of Adap1 normalized to Gapdh in the respective cell lysates ( n = 4 independent cell isolations). Full-length blots are presented in Supplementary Fig. 1 .
    Figure Legend Snippet: Adap1 expression in cardiac cells. ( A ) Relative levels of Adap1 mRNA expression in the rat brain (adult) and heart (adult and 2-day-old neonate) were measured by RT-qPCR and were normalized to the Rpl30 reporter gene ( n = 4 independent tissues). ( B ) Representative Western blots of Adap1 and Gapdh (loading control) detected from whole brain and heart extracts. ( C ) The histogram represents the relative expression level of Adap1 normalized to Gapdh in the respective tissues ( n = 4 independent tissues). ( D ) Representative Western blot of Adap1, α-Actinin (cardiomyocyte specific marker), and Gapdh (loading control) detected in the protein lysates of enriched rat neonatal ventricular cardiomyocytes (RNVC) and non-cardiomyocytes (Non-CM). ( E ) The histogram represents the relative expression level of Adap1 normalized to Gapdh in the respective cell lysates ( n = 4 independent cell isolations). Full-length blots are presented in Supplementary Fig. 1 .

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Marker

    ADAP1 reduces cell surface β1-integrin expression in cardiomyocytes. ( A ) Representative Western blot of biotin-labeled cell surface β1-integrin expression compared to total β1-integrin expression in protein lysates of rat neonatal ventricular cardiomyocytes (RNVC) infected with a β-Gal- (negative control), ADAP1-, or Mek1ca-overexpressing adenovirus, individually or in combination as indicated, and cultured for 72 h post-infection. The expression levels of ADAP1, Mek1ca, and Gapdh (loading control) were also verified. ( B ) The histogram represents the level of cell surface β1-integrin expression normalized to total β1-integrin expression relative to the β-Gal control and measured using the same experimental conditions as in A ( n = 3 independent experiments). * P
    Figure Legend Snippet: ADAP1 reduces cell surface β1-integrin expression in cardiomyocytes. ( A ) Representative Western blot of biotin-labeled cell surface β1-integrin expression compared to total β1-integrin expression in protein lysates of rat neonatal ventricular cardiomyocytes (RNVC) infected with a β-Gal- (negative control), ADAP1-, or Mek1ca-overexpressing adenovirus, individually or in combination as indicated, and cultured for 72 h post-infection. The expression levels of ADAP1, Mek1ca, and Gapdh (loading control) were also verified. ( B ) The histogram represents the level of cell surface β1-integrin expression normalized to total β1-integrin expression relative to the β-Gal control and measured using the same experimental conditions as in A ( n = 3 independent experiments). * P

    Techniques Used: Expressing, Western Blot, Labeling, Infection, Negative Control, Cell Culture

    26) Product Images from "18F-labeled Rhodamines as Potential Myocardial Perfusion Agents: Comparison of Pharmacokinetic Properties of Several Rhodamines"

    Article Title: 18F-labeled Rhodamines as Potential Myocardial Perfusion Agents: Comparison of Pharmacokinetic Properties of Several Rhodamines

    Journal: Nuclear medicine and biology

    doi: 10.1016/j.nucmedbio.2015.06.008

    Fluorescence microscopy images of rat cardiomyocytes showing mitochondria stained by MitoTracker green dye (A), [ 18 F]4 (B), nuclei stained by DAPI (C), and the overlay (D). The tracer [ 18 F]4 clearly co-localizes in the mitochondria of rat cardiomyocytes (Pearson correlation coefficient 0.85).
    Figure Legend Snippet: Fluorescence microscopy images of rat cardiomyocytes showing mitochondria stained by MitoTracker green dye (A), [ 18 F]4 (B), nuclei stained by DAPI (C), and the overlay (D). The tracer [ 18 F]4 clearly co-localizes in the mitochondria of rat cardiomyocytes (Pearson correlation coefficient 0.85).

    Techniques Used: Fluorescence, Microscopy, Staining

    Time-dependent uptake of [ 18 F]4 into rat cardiomyocytes. Error bars indicate estimated standard deviation.
    Figure Legend Snippet: Time-dependent uptake of [ 18 F]4 into rat cardiomyocytes. Error bars indicate estimated standard deviation.

    Techniques Used: Standard Deviation

    27) Product Images from "Delivery of an engineered HGF fragment in an extracellular matrix-derived hydrogel prevents negative LV remodeling post-myocardial infarction"

    Article Title: Delivery of an engineered HGF fragment in an extracellular matrix-derived hydrogel prevents negative LV remodeling post-myocardial infarction

    Journal: Biomaterials

    doi: 10.1016/j.biomaterials.2014.12.021

    Interstitial fibrosis and cardiomyocyte cross-sectional area
    Figure Legend Snippet: Interstitial fibrosis and cardiomyocyte cross-sectional area

    Techniques Used:

    28) Product Images from "Angiotensin II activates CaV1.2 Ca2+ channels through β‐arrestin2 and casein kinase 2 in mouse immature cardiomyocytes"

    Article Title: Angiotensin II activates CaV1.2 Ca2+ channels through β‐arrestin2 and casein kinase 2 in mouse immature cardiomyocytes

    Journal: The Journal of Physiology

    doi: 10.1113/JP273883

    A signaling pathway revealed by this study Schematic illustration of the AngII‐mediated activation of Ca V 1.2 channels in immature cardiomyocytes.
    Figure Legend Snippet: A signaling pathway revealed by this study Schematic illustration of the AngII‐mediated activation of Ca V 1.2 channels in immature cardiomyocytes.

    Techniques Used: Activation Assay

    Expression levels of CK2α, α’ and β during the development of mouse ventricular cardiomyocytes A and B , mouse VCMs [embryonic days (E) 14.5 from 3 pregnant mice, E19.5 from 3 pregnant mice, P0 from 5 pups in each sample, P10 from 3 mice, P28 from 3 mice and P56 from 3 mice] were extracted. These whole‐cell lysates (10 μg per lane) were immunoblotted with indicated antibodies. Data were normalized to each P0 group; n = 3 independent blots. * P
    Figure Legend Snippet: Expression levels of CK2α, α’ and β during the development of mouse ventricular cardiomyocytes A and B , mouse VCMs [embryonic days (E) 14.5 from 3 pregnant mice, E19.5 from 3 pregnant mice, P0 from 5 pups in each sample, P10 from 3 mice, P28 from 3 mice and P56 from 3 mice] were extracted. These whole‐cell lysates (10 μg per lane) were immunoblotted with indicated antibodies. Data were normalized to each P0 group; n = 3 independent blots. * P

    Techniques Used: Expressing, Mouse Assay

    29) Product Images from "Givinostat reduces adverse cardiac remodeling through regulating fibroblasts activation"

    Article Title: Givinostat reduces adverse cardiac remodeling through regulating fibroblasts activation

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-017-0174-5

    Histological analysis. a Apoptotic cardiomyocytes (α-SARC, magenta) were detected by TUNEL labeling (yellow) in SHAM and AMI mice at day 7. Scale bar represent 65 μm. The graph illustrates the ratio between the number of double positive cells (α-SARC + and TUNEL + cells) and the nuclei at different time points. b Cross sectional area (CSA) in the border zone at day 30 stained with wheat germ agglutinin (WGA, yellow). Scale bar represent 50 μm. Plot indicates myocytes CSA (μm 2 ) in both the border and the contralateral zone at different time points. c Apoptotic cells were detected by TUNEL labeling (yellow) at day 7. Scale bar represent 100 μm. The graph indicates the ratio between the number of whole TUNEL positive cells and the nuclei at different time points. d α-SMA positive vessels labeled in magenta after 30 days. Scale bar represent 100 μm. The corresponding diagram shows the ratio number of α-SMA positive vessels on total area (mm 2 ) at different time points. e Isolectin B4 positive capillaries (yellow) in the border zone at day 30. Scale bar represent 100 μm. Graphs explain the proportion number of ISO/B4 positive capillaries/cell number in the border and the contralateral zone at different time points. f Staining for Cx43 (yellow) in the border zone of the infarcted area at day 30. Scale bar represent 100 μm. Cx43 quantification, expressed as pixel 2 /total area, is elucidated in the side chart. For Cx43 quantification method details see Materials and methods section and supplementary Figure S5 . g Heart cross section of vessel composed of smooth muscle cells (α-SMA, magenta) and apoptotic cells (yellow) highlighted by arrows (white). Scale bar represent 25 μm. h Western blot analysis for Cx43 in AMI Givinostat mice vs. AMI Saline group at different time points. N = 6 mice for time point. Error bars represent ± SEM. Student’s t test, * p
    Figure Legend Snippet: Histological analysis. a Apoptotic cardiomyocytes (α-SARC, magenta) were detected by TUNEL labeling (yellow) in SHAM and AMI mice at day 7. Scale bar represent 65 μm. The graph illustrates the ratio between the number of double positive cells (α-SARC + and TUNEL + cells) and the nuclei at different time points. b Cross sectional area (CSA) in the border zone at day 30 stained with wheat germ agglutinin (WGA, yellow). Scale bar represent 50 μm. Plot indicates myocytes CSA (μm 2 ) in both the border and the contralateral zone at different time points. c Apoptotic cells were detected by TUNEL labeling (yellow) at day 7. Scale bar represent 100 μm. The graph indicates the ratio between the number of whole TUNEL positive cells and the nuclei at different time points. d α-SMA positive vessels labeled in magenta after 30 days. Scale bar represent 100 μm. The corresponding diagram shows the ratio number of α-SMA positive vessels on total area (mm 2 ) at different time points. e Isolectin B4 positive capillaries (yellow) in the border zone at day 30. Scale bar represent 100 μm. Graphs explain the proportion number of ISO/B4 positive capillaries/cell number in the border and the contralateral zone at different time points. f Staining for Cx43 (yellow) in the border zone of the infarcted area at day 30. Scale bar represent 100 μm. Cx43 quantification, expressed as pixel 2 /total area, is elucidated in the side chart. For Cx43 quantification method details see Materials and methods section and supplementary Figure S5 . g Heart cross section of vessel composed of smooth muscle cells (α-SMA, magenta) and apoptotic cells (yellow) highlighted by arrows (white). Scale bar represent 25 μm. h Western blot analysis for Cx43 in AMI Givinostat mice vs. AMI Saline group at different time points. N = 6 mice for time point. Error bars represent ± SEM. Student’s t test, * p

    Techniques Used: TUNEL Assay, Labeling, Mouse Assay, Staining, Whole Genome Amplification, Western Blot

    30) Product Images from "Innate lymphotoxin receptor mediated signaling promotes HSV-1 associated neuroinflammation and viral replication"

    Article Title: Innate lymphotoxin receptor mediated signaling promotes HSV-1 associated neuroinflammation and viral replication

    Journal: Scientific Reports

    doi: 10.1038/srep10406

    Blockade of LT/LIGHT signaling inhibits viral replication in nervous tissue. ( a ) Rag1 –/– mice (n = 4 to 7/group) were infected with 2 × 10 6 pfu of HSV-1 and treated with either LTβR-Ig or control protein on day -1 and day 5 p.i.. At the indicated time points of figures, mice were euthanized. Footpad, DRG (L3, L4 and L5) and spinal cord were collected. Viral loads in different tissue homogenates were determined by plaque assay. ( b ) Rag1 –/– mice (n = 4 /group) were infected with 2 × 10 6 pfu of HSV-1 via footpad injection. For group with sciatic nerve (SN) resection, one segment of the SN was removed on day 2 (d2) p.i., For the group with SN resection and blockade of LT/LIGHT, 100 ug/mice of LTβR-Ig was administrated on day 2 p.i. (after sciatic nerve resection) and day 8 p.i. Data are representative of two independent experiments. Statistical analysis for a. unpaired t test, Error bar represents SEM, *p
    Figure Legend Snippet: Blockade of LT/LIGHT signaling inhibits viral replication in nervous tissue. ( a ) Rag1 –/– mice (n = 4 to 7/group) were infected with 2 × 10 6 pfu of HSV-1 and treated with either LTβR-Ig or control protein on day -1 and day 5 p.i.. At the indicated time points of figures, mice were euthanized. Footpad, DRG (L3, L4 and L5) and spinal cord were collected. Viral loads in different tissue homogenates were determined by plaque assay. ( b ) Rag1 –/– mice (n = 4 /group) were infected with 2 × 10 6 pfu of HSV-1 via footpad injection. For group with sciatic nerve (SN) resection, one segment of the SN was removed on day 2 (d2) p.i., For the group with SN resection and blockade of LT/LIGHT, 100 ug/mice of LTβR-Ig was administrated on day 2 p.i. (after sciatic nerve resection) and day 8 p.i. Data are representative of two independent experiments. Statistical analysis for a. unpaired t test, Error bar represents SEM, *p

    Techniques Used: Mouse Assay, Infection, Plaque Assay, Injection

    Blockade of LT/LIGHT inhibits chemokine expression and inflammatory cell infiltration into the DRG of infected Rag1 –/– mice. Rag1 –/– mice (n = 3 to 5/group) were infected with HSV-1 and treated with LTβR-Ig as Fig. 1a . Uninfected mice were chosen as the control group. On day 6 p.i., DRGs (L3, L4 and L5) were collected from euthanized mice. The mRNA level of various chemokines (CCL2, CXCL10 and CXCL1/2) ( a ) and viral ICP0 gene ( b ) were measured by real-time PCR. Data are representative of two independent experiments. ( c ) On day 8 p.i., innate immune cell subsets in DRG were determined by flow cytometry assay. Gate strategy: monocytes (CD45 + CD11b + Ly6C hi Ly6G middle ), macrophage (CD45 + F4/80 + ). Data are pooled from two independent experiments, n = 5 to 6/group. Statistical analysis for a , b , c was by unpaired t test. Error bar represents SEM, *p
    Figure Legend Snippet: Blockade of LT/LIGHT inhibits chemokine expression and inflammatory cell infiltration into the DRG of infected Rag1 –/– mice. Rag1 –/– mice (n = 3 to 5/group) were infected with HSV-1 and treated with LTβR-Ig as Fig. 1a . Uninfected mice were chosen as the control group. On day 6 p.i., DRGs (L3, L4 and L5) were collected from euthanized mice. The mRNA level of various chemokines (CCL2, CXCL10 and CXCL1/2) ( a ) and viral ICP0 gene ( b ) were measured by real-time PCR. Data are representative of two independent experiments. ( c ) On day 8 p.i., innate immune cell subsets in DRG were determined by flow cytometry assay. Gate strategy: monocytes (CD45 + CD11b + Ly6C hi Ly6G middle ), macrophage (CD45 + F4/80 + ). Data are pooled from two independent experiments, n = 5 to 6/group. Statistical analysis for a , b , c was by unpaired t test. Error bar represents SEM, *p

    Techniques Used: Expressing, Infection, Mouse Assay, Real-time Polymerase Chain Reaction, Flow Cytometry, Cytometry

    31) Product Images from "Conditional Dicer1 depletion using Chrnb4-Cre leads to cone cell death and impaired photopic vision"

    Article Title: Conditional Dicer1 depletion using Chrnb4-Cre leads to cone cell death and impaired photopic vision

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-38294-9

    Chrnb4-GFP and Chrnb4-cre expression in developing and adult cone photoreceptor cells. (A) Immunostaining of adult (6 week old) Chrnb4-GFP retinas. Chrnb4-GFP expression co-labels with the expression of cone marker CA in the ONL. (B–D) Immunostaining of P8 Chrnb4-GFP retinas. Chrnb4-GFP expression co-labels with the expression of cone markers RxRγ (B) and CA (C) but not with the expression of rod marker RHO (D) . See Supplementary Figs 3 and 4 for the separate channels of the data shown in the merged images in A B. (E , F) Immunostaining on E17 Chrnb4-cre; R26 YFP retinas using an anti-GFP antibody to amplify the YFP signal intensity. YFP expression is detected in some RxRγ +ve cones (white arrows), while most cones remain YFP -ve ( E , yellow arrows). YFP expression is also seen in some OTX2-expressing photoreceptor progenitors ( F , white arrows). Yellow arrows indicate OTX2 +ve cells that are not YFP +ve . OTX2 +ve RPE cells do not express YFP (blue arrows). (G – I) Immunostaining on adult (6 weeks old) Chrnb4-cre; R26 YFP retinas. YFP expression was detected in CA +ve cones ( G , white arrows) and in a few inner retinal cells including some PAX6 +ve cells ( H , white arrowheads). Most Pax6 +ve cells in the inner retina remained YFP -ve ( H , yellow arrowheads). CHX10 +ve bipolar cells were mainly YFP -ve ( I , asterisks). (J) Immunostaining on E12 Chrnb4-cre; R26 YFP retinas was also performed. YFP expression was detected in a subset of CHX10 +ve RPCs (white arrows). CA: cone arrestin. RxRγ: retinoid x receptor gamma. RHO: rhodopsin. RPE: retinal pigment epithelium. ONL: outer nuclear layer. INL: inner nuclear layer. GCL: ganglion cell layer. RPC: retinal progenitor cells. Scale bars: 30 μm. Scale bars of insets: 10 μm.
    Figure Legend Snippet: Chrnb4-GFP and Chrnb4-cre expression in developing and adult cone photoreceptor cells. (A) Immunostaining of adult (6 week old) Chrnb4-GFP retinas. Chrnb4-GFP expression co-labels with the expression of cone marker CA in the ONL. (B–D) Immunostaining of P8 Chrnb4-GFP retinas. Chrnb4-GFP expression co-labels with the expression of cone markers RxRγ (B) and CA (C) but not with the expression of rod marker RHO (D) . See Supplementary Figs 3 and 4 for the separate channels of the data shown in the merged images in A B. (E , F) Immunostaining on E17 Chrnb4-cre; R26 YFP retinas using an anti-GFP antibody to amplify the YFP signal intensity. YFP expression is detected in some RxRγ +ve cones (white arrows), while most cones remain YFP -ve ( E , yellow arrows). YFP expression is also seen in some OTX2-expressing photoreceptor progenitors ( F , white arrows). Yellow arrows indicate OTX2 +ve cells that are not YFP +ve . OTX2 +ve RPE cells do not express YFP (blue arrows). (G – I) Immunostaining on adult (6 weeks old) Chrnb4-cre; R26 YFP retinas. YFP expression was detected in CA +ve cones ( G , white arrows) and in a few inner retinal cells including some PAX6 +ve cells ( H , white arrowheads). Most Pax6 +ve cells in the inner retina remained YFP -ve ( H , yellow arrowheads). CHX10 +ve bipolar cells were mainly YFP -ve ( I , asterisks). (J) Immunostaining on E12 Chrnb4-cre; R26 YFP retinas was also performed. YFP expression was detected in a subset of CHX10 +ve RPCs (white arrows). CA: cone arrestin. RxRγ: retinoid x receptor gamma. RHO: rhodopsin. RPE: retinal pigment epithelium. ONL: outer nuclear layer. INL: inner nuclear layer. GCL: ganglion cell layer. RPC: retinal progenitor cells. Scale bars: 30 μm. Scale bars of insets: 10 μm.

    Techniques Used: Expressing, Immunostaining, Marker

    32) Product Images from "Liver X receptors are negative regulators of cardiac hypertrophy via suppressing NF-?B signalling"

    Article Title: Liver X receptors are negative regulators of cardiac hypertrophy via suppressing NF-?B signalling

    Journal: Cardiovascular Research

    doi: 10.1093/cvr/cvp180

    Effect of LXR agonist T1317 on AngII-induced cardiac hypertrophy in RNCM. RNCM were stimulated with 1 µM AngII in the presence or absence of T1317 (1, 5, and 10 µmol/L) that was added 2 h before experiments. ( A ) Effects of AngII with and without T1317 on cell surface area of cardiomyocytes. Cell surface area was calculated by measuring 50 random cardiomyocytes with Image Pro. ( n = 6, * P
    Figure Legend Snippet: Effect of LXR agonist T1317 on AngII-induced cardiac hypertrophy in RNCM. RNCM were stimulated with 1 µM AngII in the presence or absence of T1317 (1, 5, and 10 µmol/L) that was added 2 h before experiments. ( A ) Effects of AngII with and without T1317 on cell surface area of cardiomyocytes. Cell surface area was calculated by measuring 50 random cardiomyocytes with Image Pro. ( n = 6, * P

    Techniques Used:

    33) Product Images from "Nuclear CaMKII enhances histone H3 phosphorylation and remodels chromatin during cardiac hypertrophy"

    Article Title: Nuclear CaMKII enhances histone H3 phosphorylation and remodels chromatin during cardiac hypertrophy

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkt500

    Increased histone H3 phosphorylation during hypertrophic stimulation. ( A ) Western analysis from primary cardiac cells stimulated with PE and XY using anti-H3 Ser-10, p-CaMKII, α-actinin and GAPDH antibodies. This experiment was repeated three times with three independent cardiomyocyte preparations. ( B ) Quantitation of A. SF: cells maintained in serum-free conditions. ( C ) Western blot analysis of cell extracts prepared from primary cardiomyocytes treated with PE and transfected with control siRNA (siCt) or siRNA against CaMKIIδB (siδB), using α-histone H3 Ser-10 antibody. NT: non-transfected cells ( n = 3). ( D ) Specific silencing of CaMKIIδB by siRNA decreases H3 Ser-10 in ventricular myocytes undergoing hypertrophy. Merge image showing decreased histone H3 phosphorylation in ventricular myocytes stimulated with PE for 24 h transfected with siCaMKIIδB compared with siControl using indirect immunofluorescence. Nuclei are shown in blue with 4′,6-diamidino-2-phenylindole (DAPI) staining, histone H3 Ser-10 is in green. Ventricular cardiomyocytes stained with α-actinin antibody are shown in red. Arrows indicate the nuclei of myocytes positive for H3 Ser-10 in siControl myocytes and arrowheads nuclei which are negative for H3 Ser-10 in ventricular myocytes after CaMKIIδB knock-down. ( E ) Percentage of ventricular myocytes positive for H3 Ser-10. Quantitative analysis of B was done in five different fields. * P
    Figure Legend Snippet: Increased histone H3 phosphorylation during hypertrophic stimulation. ( A ) Western analysis from primary cardiac cells stimulated with PE and XY using anti-H3 Ser-10, p-CaMKII, α-actinin and GAPDH antibodies. This experiment was repeated three times with three independent cardiomyocyte preparations. ( B ) Quantitation of A. SF: cells maintained in serum-free conditions. ( C ) Western blot analysis of cell extracts prepared from primary cardiomyocytes treated with PE and transfected with control siRNA (siCt) or siRNA against CaMKIIδB (siδB), using α-histone H3 Ser-10 antibody. NT: non-transfected cells ( n = 3). ( D ) Specific silencing of CaMKIIδB by siRNA decreases H3 Ser-10 in ventricular myocytes undergoing hypertrophy. Merge image showing decreased histone H3 phosphorylation in ventricular myocytes stimulated with PE for 24 h transfected with siCaMKIIδB compared with siControl using indirect immunofluorescence. Nuclei are shown in blue with 4′,6-diamidino-2-phenylindole (DAPI) staining, histone H3 Ser-10 is in green. Ventricular cardiomyocytes stained with α-actinin antibody are shown in red. Arrows indicate the nuclei of myocytes positive for H3 Ser-10 in siControl myocytes and arrowheads nuclei which are negative for H3 Ser-10 in ventricular myocytes after CaMKIIδB knock-down. ( E ) Percentage of ventricular myocytes positive for H3 Ser-10. Quantitative analysis of B was done in five different fields. * P

    Techniques Used: Western Blot, Quantitation Assay, Transfection, Immunofluorescence, Staining

    CaMKIIδB binds to cardiac chromatin in vitro and in primary cardiomyocytes. ( A ) Coomassie staining of proteins that interact with inactive HA-tagged CaMKIIδB-K43A in cardiac cells, after immunoprecipitation of nuclear extracts using HA agarose affinity gels and analysis by SDS–PAGE. Arrows indicate CaMKIIδB and the histone proteins later identified by mass spectrometry that co-migrate with purified histone octamers. Asterisks show immunoprecipitated proteins at 150 mM salt concentration in cells expressing CaMKIIδB-K43A or GFP. ( B ) Western blot analysis showing interaction of CaMKIIδB-K43A with histone proteins after immunoprecipitation with HA affinity gels ( n = 3). Purified histone octamers were run in parallel as a positive control. ( C ) Interaction of endogenous histone proteins with endogenous CaMKIIδB shown after immunoprecipitation of cardiac nuclear extracts with α-H3 or control IgG antibodies and immunoblotting using a CaMKIIδ-specific antibody. ( D ) Reverse experiment where CaMKIIδ was immunoprecipitated first with anti-CaMKIIδ antibody, and the precipitated proteins were analyzed by SDS–PAGE and immunoblotted with α-H3-specific antibody. Blots in C and D are representative of two independent experiments. ( E ) Western blot analysis showing the fraction of cardio-chromatin that binds to CaMKIIδB attached to HA agarose or that remains in the supernatant ( n = 4). Immunoblotting with anti-H2B and -HA antibodies from 1: input chromatin, 2: chromatin bound to HA-CaMKIIδB, 3: chromatin remaining in the supernatant not bound to HA-CaMKIIδB, 4: chromatin bound to HA beads alone, 5: chromatin remaining in the supernatant not attached to HA beads. ( F ) Representative EMSA with increasing concentration of purified CaMKIIδB and 0W0 or 0W47 mononucleosome templates followed by native PAGE, n = 3. Arrows indicates the position of ISWI or CaMKIIδB bound to mononucleosomes.
    Figure Legend Snippet: CaMKIIδB binds to cardiac chromatin in vitro and in primary cardiomyocytes. ( A ) Coomassie staining of proteins that interact with inactive HA-tagged CaMKIIδB-K43A in cardiac cells, after immunoprecipitation of nuclear extracts using HA agarose affinity gels and analysis by SDS–PAGE. Arrows indicate CaMKIIδB and the histone proteins later identified by mass spectrometry that co-migrate with purified histone octamers. Asterisks show immunoprecipitated proteins at 150 mM salt concentration in cells expressing CaMKIIδB-K43A or GFP. ( B ) Western blot analysis showing interaction of CaMKIIδB-K43A with histone proteins after immunoprecipitation with HA affinity gels ( n = 3). Purified histone octamers were run in parallel as a positive control. ( C ) Interaction of endogenous histone proteins with endogenous CaMKIIδB shown after immunoprecipitation of cardiac nuclear extracts with α-H3 or control IgG antibodies and immunoblotting using a CaMKIIδ-specific antibody. ( D ) Reverse experiment where CaMKIIδ was immunoprecipitated first with anti-CaMKIIδ antibody, and the precipitated proteins were analyzed by SDS–PAGE and immunoblotted with α-H3-specific antibody. Blots in C and D are representative of two independent experiments. ( E ) Western blot analysis showing the fraction of cardio-chromatin that binds to CaMKIIδB attached to HA agarose or that remains in the supernatant ( n = 4). Immunoblotting with anti-H2B and -HA antibodies from 1: input chromatin, 2: chromatin bound to HA-CaMKIIδB, 3: chromatin remaining in the supernatant not bound to HA-CaMKIIδB, 4: chromatin bound to HA beads alone, 5: chromatin remaining in the supernatant not attached to HA beads. ( F ) Representative EMSA with increasing concentration of purified CaMKIIδB and 0W0 or 0W47 mononucleosome templates followed by native PAGE, n = 3. Arrows indicates the position of ISWI or CaMKIIδB bound to mononucleosomes.

    Techniques Used: In Vitro, Staining, Immunoprecipitation, SDS Page, Mass Spectrometry, Purification, Concentration Assay, Expressing, Western Blot, Positive Control, Clear Native PAGE

    H3 Ser-10 phosphorylation in ventricular myocytes is not associated with cellular proliferation. ( A ) Expression level of cell cycle proteins and cycle-dependent kinases during hypertrophic stimulation. Immunoblots from primary cardiac cells stimulated with PE and XY using anti-Cdk4, cyclin E, Cdc2 and GAPDH antibodies. This experiment was repeated twice in two independent cardiomyocyte preparations. SF: serum-free conditions. ( B ) Immunocytochemistry for BrdU and immunofluorescence for Ki67 in primary neonatal rat cardiomyocytes treated with PE for 24 h after transfection with siControl or siCaMKIIδB. CS: cardiomyocytes treated with serum. For Ki67, nuclei are shown in blue with DAPI staining, Ki67 is shown in red. Ventricular cardiomyocytes stained with α-actinin antibody are shown in green.
    Figure Legend Snippet: H3 Ser-10 phosphorylation in ventricular myocytes is not associated with cellular proliferation. ( A ) Expression level of cell cycle proteins and cycle-dependent kinases during hypertrophic stimulation. Immunoblots from primary cardiac cells stimulated with PE and XY using anti-Cdk4, cyclin E, Cdc2 and GAPDH antibodies. This experiment was repeated twice in two independent cardiomyocyte preparations. SF: serum-free conditions. ( B ) Immunocytochemistry for BrdU and immunofluorescence for Ki67 in primary neonatal rat cardiomyocytes treated with PE for 24 h after transfection with siControl or siCaMKIIδB. CS: cardiomyocytes treated with serum. For Ki67, nuclei are shown in blue with DAPI staining, Ki67 is shown in red. Ventricular cardiomyocytes stained with α-actinin antibody are shown in green.

    Techniques Used: Expressing, Western Blot, Immunocytochemistry, Immunofluorescence, Transfection, Staining

    Increased histone H3 Ser-10 and CaMKIIδB recruitment at fetal-cardiac promoters during hypertrophic stimulation. ( A ) Primers used for the amplification of hypertrophic genes after ChIP from primary cardiomyocytes stimulated with PE. Primer sets are specific for the ANF, β-MHC, α-cardiac actin and GAPDH genes. Black boxes represent the position of the primers relative to the start of transcription. ChIP–Q-PCR assay using ( B ) α-histone H3 Ser-10 or ( C ) CaMKIIδ antibodies to precipitate chromatin from primary neonatal rat cardiomyocytes maintained in serum-free conditions or treated with PE for 6, 24 and 24 h. Results are expressed as % input over basal condition (serum-free treatment). Error bars represent means ± SD ( n = 3). ** P
    Figure Legend Snippet: Increased histone H3 Ser-10 and CaMKIIδB recruitment at fetal-cardiac promoters during hypertrophic stimulation. ( A ) Primers used for the amplification of hypertrophic genes after ChIP from primary cardiomyocytes stimulated with PE. Primer sets are specific for the ANF, β-MHC, α-cardiac actin and GAPDH genes. Black boxes represent the position of the primers relative to the start of transcription. ChIP–Q-PCR assay using ( B ) α-histone H3 Ser-10 or ( C ) CaMKIIδ antibodies to precipitate chromatin from primary neonatal rat cardiomyocytes maintained in serum-free conditions or treated with PE for 6, 24 and 24 h. Results are expressed as % input over basal condition (serum-free treatment). Error bars represent means ± SD ( n = 3). ** P

    Techniques Used: Amplification, Chromatin Immunoprecipitation, Polymerase Chain Reaction

    CaMKIIδB specifically phosphorylates histone H3 in histone octamers and in chromatin. ( A ) In vitro kinase assay ( n = 3) with purified constitutively active CaMKIIδB-T287D and reconstituted histone octamers in the presence of [γ 32 P] ATP followed by SDS–PAGE and autoradiography. Asterisks indicate phosphorylation of histone H3 by CaMKIIδB. ( B ) In vitro kinase assays with constitutively active CaMKIIδB-T287D and de-phosphorylated chromatin after treatment with calf intestine alkaline phosphatase (CIAP), purified from neonatal rat cardiomyocytes and HeLa cells in the presence of [γ 32 P] ATP followed by SDS–PAGE ( n = 5). Asterisks indicate phosphorylated histone H3. Lane 1: input de-phosphorylated cardio-chromatin, 2: input de-phosphorylated HeLa chromatin, 3: CaMKIIδB-T287D alone, 4: CaMKIIδB-T287D and de-phosphorylated cardio-chromatin, 5: CaMKIIδB-T287D and de-phosphorylated HeLa chromatin. ( C ) Western blot analysis with phospho-H3 (Ser-10), phospho-H3 (Ser-28) or total H3-specific antibodies after in vitro kinase assay with recombinant histone H3 and active CaMKIIδB-T287D ( n = 3). ( D ) Immunoblots after in vitro phosphorylation assay with endogenous cardio-chromatin or chromatin de-phosphorylated by CIAP treatment in the presence of increasing amount of CaMKIIδB-T287D enzyme, using anti-H3 Ser-10, -H2B and -HA antibodies ( n = 3). Lane 1: cardio-chromatin alone, 2: cardio-chromatin treated with CIAP, 3–5: cardio-chromatin treated with CIAP incubated with different amounts of CaMKIIδB-T287D. ( E ) In vitro kinase assay in the presence of [γ 32 P] ATP after transfection of HeLa cells with CaMKIIδB-T287D and wild-type Flag-histone H3 or mutant histone H3 S10A and immunoprecipitation of isolated chromatin using Flag resins ( n = 3). Lane 1: Flag-H3 wild-type alone, 2: Flag-H3-S10A alone, 3: CaMKIIδB-T287D alone, 4: Flag-H3 wild-type + CaMKIIδB-T287D, 5: Flag-H3-S10A + CaMKIIδB-T287D. ( F ) Immunoblots showing phosphorylation at Ser-10 of wild-type Flag-histone H3 and three different H3 S10A mutants after transfection in HeLa cells and immunoprecipitation using Flag resins ( n = 3). Lane 1: Flag H3 wild-type, 2: Flag-H3 wild-type treated with CIAP, 3–5: Flag-H3-S10A from three separate clones.
    Figure Legend Snippet: CaMKIIδB specifically phosphorylates histone H3 in histone octamers and in chromatin. ( A ) In vitro kinase assay ( n = 3) with purified constitutively active CaMKIIδB-T287D and reconstituted histone octamers in the presence of [γ 32 P] ATP followed by SDS–PAGE and autoradiography. Asterisks indicate phosphorylation of histone H3 by CaMKIIδB. ( B ) In vitro kinase assays with constitutively active CaMKIIδB-T287D and de-phosphorylated chromatin after treatment with calf intestine alkaline phosphatase (CIAP), purified from neonatal rat cardiomyocytes and HeLa cells in the presence of [γ 32 P] ATP followed by SDS–PAGE ( n = 5). Asterisks indicate phosphorylated histone H3. Lane 1: input de-phosphorylated cardio-chromatin, 2: input de-phosphorylated HeLa chromatin, 3: CaMKIIδB-T287D alone, 4: CaMKIIδB-T287D and de-phosphorylated cardio-chromatin, 5: CaMKIIδB-T287D and de-phosphorylated HeLa chromatin. ( C ) Western blot analysis with phospho-H3 (Ser-10), phospho-H3 (Ser-28) or total H3-specific antibodies after in vitro kinase assay with recombinant histone H3 and active CaMKIIδB-T287D ( n = 3). ( D ) Immunoblots after in vitro phosphorylation assay with endogenous cardio-chromatin or chromatin de-phosphorylated by CIAP treatment in the presence of increasing amount of CaMKIIδB-T287D enzyme, using anti-H3 Ser-10, -H2B and -HA antibodies ( n = 3). Lane 1: cardio-chromatin alone, 2: cardio-chromatin treated with CIAP, 3–5: cardio-chromatin treated with CIAP incubated with different amounts of CaMKIIδB-T287D. ( E ) In vitro kinase assay in the presence of [γ 32 P] ATP after transfection of HeLa cells with CaMKIIδB-T287D and wild-type Flag-histone H3 or mutant histone H3 S10A and immunoprecipitation of isolated chromatin using Flag resins ( n = 3). Lane 1: Flag-H3 wild-type alone, 2: Flag-H3-S10A alone, 3: CaMKIIδB-T287D alone, 4: Flag-H3 wild-type + CaMKIIδB-T287D, 5: Flag-H3-S10A + CaMKIIδB-T287D. ( F ) Immunoblots showing phosphorylation at Ser-10 of wild-type Flag-histone H3 and three different H3 S10A mutants after transfection in HeLa cells and immunoprecipitation using Flag resins ( n = 3). Lane 1: Flag H3 wild-type, 2: Flag-H3 wild-type treated with CIAP, 3–5: Flag-H3-S10A from three separate clones.

    Techniques Used: In Vitro, Kinase Assay, Purification, SDS Page, Autoradiography, Western Blot, Recombinant, Phosphorylation Assay, Incubation, Transfection, Mutagenesis, Immunoprecipitation, Isolation, Clone Assay

    34) Product Images from "Tsg101 positively regulates physiologic-like cardiac hypertrophy through FIP3-mediated endosomal recycling of IGF-1R"

    Article Title: Tsg101 positively regulates physiologic-like cardiac hypertrophy through FIP3-mediated endosomal recycling of IGF-1R

    Journal: The FASEB Journal

    doi: 10.1096/fj.201802338RR

    Overexpression of Tsg101 enhances recycling of IGF-1R in cardiomyocytes. A , B ) Representative immunoblots ( A ) and quantitative analyses ( B ) of streptavidin immunoprecipitates (IP), visualized using an IGF-1R antibody. Whole-cell lysates (WCLs) were used as loading controls. Four independent experiments were performed for recycling assays. C ) Representative images of immunofluorescence staining with streptavidin Alexa Fluor 594 conjugate after IGF-1R biotin was internalized and recycled in Ad.Tsg101 and Ad.GFP NRCMs; n = 4 plates, 25–30 cells per plate. Scale bars, 10 μm. * P
    Figure Legend Snippet: Overexpression of Tsg101 enhances recycling of IGF-1R in cardiomyocytes. A , B ) Representative immunoblots ( A ) and quantitative analyses ( B ) of streptavidin immunoprecipitates (IP), visualized using an IGF-1R antibody. Whole-cell lysates (WCLs) were used as loading controls. Four independent experiments were performed for recycling assays. C ) Representative images of immunofluorescence staining with streptavidin Alexa Fluor 594 conjugate after IGF-1R biotin was internalized and recycled in Ad.Tsg101 and Ad.GFP NRCMs; n = 4 plates, 25–30 cells per plate. Scale bars, 10 μm. * P

    Techniques Used: Over Expression, Western Blot, Immunofluorescence, Staining

    Overexpression of Tsg101 in NRCMs promotes cell hypertrophy. A ) Diagrams depicting the recombinant adenoviral vectors Ad.GFP and Ad.Tsg101. B ) Representative images showing Ad.GFP- or Ad.Tsg101-infected NRCMs (left panel is under bright field; right panel is the same field under fluorescence microscope). Scale bars, 20 μm. C ) Western blot showing increased expression of Tsg101 in Ad.Tsg101 cardiomyocytes compared with Ad.GFP cells. D , E ) Measurement of myocyte cross-sectional area in those infected cells (green) that were stained with cardiomyocyte-specific α-actinin antibody (red). Scale bars, 20 μm. n = 6 plates, 25–30 cells per plate. F , G ) Representative immunoblots ( F ) and quantitative analyses ( G ) showing expression of PM IGF-1R, total IGF-1R, Akt phosphorylation at Ser473, Rab5a, Rab4a, Rab7, Rab11a, and FIP3 in Ad.Tsg101 and Ad.GFP cells. GAPDH was used as loading control for total protein, total Akt was loading control for Akt phosphorylation, and Na/K-ATPase was loading control for PM protein. PM, plasma membrane; n = 4 for independent experiments. * P
    Figure Legend Snippet: Overexpression of Tsg101 in NRCMs promotes cell hypertrophy. A ) Diagrams depicting the recombinant adenoviral vectors Ad.GFP and Ad.Tsg101. B ) Representative images showing Ad.GFP- or Ad.Tsg101-infected NRCMs (left panel is under bright field; right panel is the same field under fluorescence microscope). Scale bars, 20 μm. C ) Western blot showing increased expression of Tsg101 in Ad.Tsg101 cardiomyocytes compared with Ad.GFP cells. D , E ) Measurement of myocyte cross-sectional area in those infected cells (green) that were stained with cardiomyocyte-specific α-actinin antibody (red). Scale bars, 20 μm. n = 6 plates, 25–30 cells per plate. F , G ) Representative immunoblots ( F ) and quantitative analyses ( G ) showing expression of PM IGF-1R, total IGF-1R, Akt phosphorylation at Ser473, Rab5a, Rab4a, Rab7, Rab11a, and FIP3 in Ad.Tsg101 and Ad.GFP cells. GAPDH was used as loading control for total protein, total Akt was loading control for Akt phosphorylation, and Na/K-ATPase was loading control for PM protein. PM, plasma membrane; n = 4 for independent experiments. * P

    Techniques Used: Over Expression, Recombinant, Infection, Fluorescence, Microscopy, Western Blot, Expressing, Staining

    Tsg101-mediated cardiomyocyte hypertrophy is attenuated by monensin and KD of FIP3. A , B ) Treatment of NRCMs with monensin (1 μM) 1 h prior to adenovirus infection resulted in reduced plasma membrane levels of IGF-1R; n = 4 for independent experiments; * P
    Figure Legend Snippet: Tsg101-mediated cardiomyocyte hypertrophy is attenuated by monensin and KD of FIP3. A , B ) Treatment of NRCMs with monensin (1 μM) 1 h prior to adenovirus infection resulted in reduced plasma membrane levels of IGF-1R; n = 4 for independent experiments; * P

    Techniques Used: Infection

    Diagram showing structural components and major players involved in the Tsg101-mediated endosomal recycling of IGF-1R. Tsg101 interacts with and enhances the function of FIP3 at the ERC. Increased activity of the ERC promotes translocation of IGF-1R to the plasma membrane, which in turn augments IGF-IR and Akt signaling in cardiac myocytes, leading to physiologic cardiac hypertrophy.
    Figure Legend Snippet: Diagram showing structural components and major players involved in the Tsg101-mediated endosomal recycling of IGF-1R. Tsg101 interacts with and enhances the function of FIP3 at the ERC. Increased activity of the ERC promotes translocation of IGF-1R to the plasma membrane, which in turn augments IGF-IR and Akt signaling in cardiac myocytes, leading to physiologic cardiac hypertrophy.

    Techniques Used: Activity Assay, Translocation Assay

    35) Product Images from "Myocardial hypertrophy in the absence of external stimuli is induced by angiogenesis in mice"

    Article Title: Myocardial hypertrophy in the absence of external stimuli is induced by angiogenesis in mice

    Journal: The Journal of Clinical Investigation

    doi: 10.1172/JCI32024

    Angiogenesis-induced hypertrophy in the setting of MI. ( A ) PR39 induction in tTA + /PR39 + mice at the time of LAD ligation resulted in reduction of the infarct size 28 days after MI ( n = 6 mice/group). ( B ) Ejection fraction (EF) assessed by pressure-volume loop analysis at 28 days after MI in induced tTA + /PR39 + and control mice. ( C and D ) Cross-sectional area of cardiomyocytes in the free (normal) LV wall myocardium in induced tTA + /PR39 + and control mice expressed as percentage of control. ( D ) Representative laminin immunostaining of LV myocardium from the 2 mouse groups. ( E ) Infarct size 28 days after MI in C57BL/6 mice exposed to Ad–hVEGF-B 167 (black bars) or control virus (Ad-RR5, white bars). Sham, gray bars. Ad–hVEGF-B 167 , n = 16; Ad-RR5, n = 12; sham, n = 5. ( F and G ) Capillary count ( F ) and capillary/myocyte ratio ( G ) in mice exposed to Ad–VEGF-B 167 or Ad-RR5 28 days after MI or in sham-operated mice. ( H and I ) Cardiomyocyte hypertrophy in the remote myocardium. Laminin staining ( H ) and quantitative analysis ( I ) of cardiomyocyte area in the normal myocardium of C57BL/6 mice 28 days after MI or a sham procedure and exposed to Ad–hVEGF-B 167 or Ad-RR5. ( J ) LV function (fractional shortening) 28 days after MI or a sham procedure in C57BL/6 mice exposed to Ad–hVEGF-B 167 or Ad-RR5. * P
    Figure Legend Snippet: Angiogenesis-induced hypertrophy in the setting of MI. ( A ) PR39 induction in tTA + /PR39 + mice at the time of LAD ligation resulted in reduction of the infarct size 28 days after MI ( n = 6 mice/group). ( B ) Ejection fraction (EF) assessed by pressure-volume loop analysis at 28 days after MI in induced tTA + /PR39 + and control mice. ( C and D ) Cross-sectional area of cardiomyocytes in the free (normal) LV wall myocardium in induced tTA + /PR39 + and control mice expressed as percentage of control. ( D ) Representative laminin immunostaining of LV myocardium from the 2 mouse groups. ( E ) Infarct size 28 days after MI in C57BL/6 mice exposed to Ad–hVEGF-B 167 (black bars) or control virus (Ad-RR5, white bars). Sham, gray bars. Ad–hVEGF-B 167 , n = 16; Ad-RR5, n = 12; sham, n = 5. ( F and G ) Capillary count ( F ) and capillary/myocyte ratio ( G ) in mice exposed to Ad–VEGF-B 167 or Ad-RR5 28 days after MI or in sham-operated mice. ( H and I ) Cardiomyocyte hypertrophy in the remote myocardium. Laminin staining ( H ) and quantitative analysis ( I ) of cardiomyocyte area in the normal myocardium of C57BL/6 mice 28 days after MI or a sham procedure and exposed to Ad–hVEGF-B 167 or Ad-RR5. ( J ) LV function (fractional shortening) 28 days after MI or a sham procedure in C57BL/6 mice exposed to Ad–hVEGF-B 167 or Ad-RR5. * P

    Techniques Used: Mouse Assay, Ligation, Immunostaining, Staining

    Myocardial hypertrophy in tTA + /PR39 + mice. ( A ) Isolated cardiomyocytes from tTA + /PR39 + mice after 6 weeks induction compared with control. Scale bar: 100 μm. ( B ) Quantification of cross-sectional area of isolated cardiomyocytes. 50 cells/ group; * P
    Figure Legend Snippet: Myocardial hypertrophy in tTA + /PR39 + mice. ( A ) Isolated cardiomyocytes from tTA + /PR39 + mice after 6 weeks induction compared with control. Scale bar: 100 μm. ( B ) Quantification of cross-sectional area of isolated cardiomyocytes. 50 cells/ group; * P

    Techniques Used: Mouse Assay, Isolation

    In vitro effects of PR39 administration. ( A ) Surface area of rat neonatal cardiomyocytes transduced with Ad-PR39 (100 MOI) or treated with PR39 (1 μM) or Ang II (1 μM) peptides for 72 hours ( n = 150 cells each). ( B ) RT-PCR analysis of hypertrophic markers ANF, β-MHC, and skeletal α-actin after 72 hours of exposure to PR39 (adenoviral vector or peptide) or Ang II. ( C ) Stimulation of protein synthesis assessed by 3 H-leucine incorporation ( n = 3 independent experiments). ( D ) Micrographs of rat neonatal cardiomyocytes transduced with Ad-PR39 (100 MOI) or treated with PR39 peptide (1 μM) or Ang II (1 μM) for 72 hours, immunostained with α-actinin sarcomeric Ab. ** P
    Figure Legend Snippet: In vitro effects of PR39 administration. ( A ) Surface area of rat neonatal cardiomyocytes transduced with Ad-PR39 (100 MOI) or treated with PR39 (1 μM) or Ang II (1 μM) peptides for 72 hours ( n = 150 cells each). ( B ) RT-PCR analysis of hypertrophic markers ANF, β-MHC, and skeletal α-actin after 72 hours of exposure to PR39 (adenoviral vector or peptide) or Ang II. ( C ) Stimulation of protein synthesis assessed by 3 H-leucine incorporation ( n = 3 independent experiments). ( D ) Micrographs of rat neonatal cardiomyocytes transduced with Ad-PR39 (100 MOI) or treated with PR39 peptide (1 μM) or Ang II (1 μM) for 72 hours, immunostained with α-actinin sarcomeric Ab. ** P

    Techniques Used: In Vitro, Transduction, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation

    36) Product Images from "Down-regulation of the M6P/IGF-II receptor increases cell proliferation and reduces apoptosis in neonatal rat cardiac myocytes"

    Article Title: Down-regulation of the M6P/IGF-II receptor increases cell proliferation and reduces apoptosis in neonatal rat cardiac myocytes

    Journal: BMC Cell Biology

    doi: 10.1186/1471-2121-5-15

    Effects of the ribozyme on proliferation of cardiac myocytes. Cells were infected with Ad-GFP/IGF2R-Rz or Ad-GFP, and then analyzed by MTT assay ( A ) and by cell counts ( B ). The data are average ± SE of at least four independent experiments (*, p
    Figure Legend Snippet: Effects of the ribozyme on proliferation of cardiac myocytes. Cells were infected with Ad-GFP/IGF2R-Rz or Ad-GFP, and then analyzed by MTT assay ( A ) and by cell counts ( B ). The data are average ± SE of at least four independent experiments (*, p

    Techniques Used: Infection, MTT Assay

    Effect of the ribozyme on the apoptosis and viability of cardiac myocytes. Cells were infected with either Ad-GFP or Ad-GFP/IGF2R-Rz and challenged with hypoxia ( A ) or TNF ( B ). Apoptotic cells were assayed using the Cell Death ELISA assay and cell viability was analyzed by MTT assay. The data are average ± SE of at least three independent experiments (*, p
    Figure Legend Snippet: Effect of the ribozyme on the apoptosis and viability of cardiac myocytes. Cells were infected with either Ad-GFP or Ad-GFP/IGF2R-Rz and challenged with hypoxia ( A ) or TNF ( B ). Apoptotic cells were assayed using the Cell Death ELISA assay and cell viability was analyzed by MTT assay. The data are average ± SE of at least three independent experiments (*, p

    Techniques Used: Infection, Enzyme-linked Immunosorbent Assay, MTT Assay

    Effect of the ribozyme on M6P/IGF2R mRNA in cardiac myocytes. ( A ) A representative RT-PCR result showing the levels of M6P/IGF2R mRNA (lanes 1 and 2) and the corresponding amounts of β-actin mRNA (lanes 3 and 4) in the ribozyme-treated myocytes (lane 1, lane 3) and in the control cells (lane 2, lane 4). ( B ) Bar chart showing percent change in M6P/IGF2R mRNA of the ribozyme-treated cells relative to the mRNA fraction in the control cells (normalized to β-actin). The data are average ± SE of at least three independent experiments (*, P
    Figure Legend Snippet: Effect of the ribozyme on M6P/IGF2R mRNA in cardiac myocytes. ( A ) A representative RT-PCR result showing the levels of M6P/IGF2R mRNA (lanes 1 and 2) and the corresponding amounts of β-actin mRNA (lanes 3 and 4) in the ribozyme-treated myocytes (lane 1, lane 3) and in the control cells (lane 2, lane 4). ( B ) Bar chart showing percent change in M6P/IGF2R mRNA of the ribozyme-treated cells relative to the mRNA fraction in the control cells (normalized to β-actin). The data are average ± SE of at least three independent experiments (*, P

    Techniques Used: Reverse Transcription Polymerase Chain Reaction

    Microphotographs showing the effect of the ribozyme on the growth of neonatal rat cardiac myocytes. Cells were infected with Ad-GFP/IGF2R-Rz or Ad-GFP. The photographs were taken directly from the culture plate under 510 nm light (upper panels) and under bright light (lower panels) using a fluorescence microscope.
    Figure Legend Snippet: Microphotographs showing the effect of the ribozyme on the growth of neonatal rat cardiac myocytes. Cells were infected with Ad-GFP/IGF2R-Rz or Ad-GFP. The photographs were taken directly from the culture plate under 510 nm light (upper panels) and under bright light (lower panels) using a fluorescence microscope.

    Techniques Used: Infection, Fluorescence, Microscopy

    Effects of the IGF2R ribozyme on the apoptosis induced by hypoxia and TNF treatment in cardiac myocytes. Cells were infected with Ad-GFP (control) or Ad-GFP/IGF2R-Rz. 72 hours post infection, cells were challenged with either hypoxia ( A ) or TNF ( B ) for one day and then cell death was examined using a fluorescence microscope. Cells were stained with Hoechst dye for nuclei and observed under 480 nm blue-fluorescent light. The bright blue spots are the nuclei of apoptotic cells.
    Figure Legend Snippet: Effects of the IGF2R ribozyme on the apoptosis induced by hypoxia and TNF treatment in cardiac myocytes. Cells were infected with Ad-GFP (control) or Ad-GFP/IGF2R-Rz. 72 hours post infection, cells were challenged with either hypoxia ( A ) or TNF ( B ) for one day and then cell death was examined using a fluorescence microscope. Cells were stained with Hoechst dye for nuclei and observed under 480 nm blue-fluorescent light. The bright blue spots are the nuclei of apoptotic cells.

    Techniques Used: Infection, Fluorescence, Microscopy, Staining

    37) Product Images from "Reduced Apaf-1 levels in cardiomyocytes engage strict regulation of apoptosis by endogenous XIAP"

    Article Title: Reduced Apaf-1 levels in cardiomyocytes engage strict regulation of apoptosis by endogenous XIAP

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.200504082

    Restoring Apaf-1 levels eliminates the strict control of XIAP and permits cytochrome c to induce apoptosis in cardiomyocytes. Rat cardiomyocytes were transfected with plasmids expressing GFP alone (Vector/GFP), Apaf-1 and GFP (Apaf-1/GFP), or procaspase-9 and GFP (caspase-9/GFP). After 24 h, the transfected cells (identifiable by GFP expression) were microinjected with 25 μg/μl of bovine or yeast cytochrome c and rhodamine dextran (Cyt c/Rhod). (A) Fluorescence photographs (for the GFP- or Rhodamine-selective channels) of representative cells taken 2 h after the cytochrome c injections. Arrows point to the GFP-positive cells that were injected with cytochrome c and rhodamine. (B) Quantitation of cell survival (2 h after the injections) for the various conditions. Data shown are the mean ± SEM of three independent experiments.
    Figure Legend Snippet: Restoring Apaf-1 levels eliminates the strict control of XIAP and permits cytochrome c to induce apoptosis in cardiomyocytes. Rat cardiomyocytes were transfected with plasmids expressing GFP alone (Vector/GFP), Apaf-1 and GFP (Apaf-1/GFP), or procaspase-9 and GFP (caspase-9/GFP). After 24 h, the transfected cells (identifiable by GFP expression) were microinjected with 25 μg/μl of bovine or yeast cytochrome c and rhodamine dextran (Cyt c/Rhod). (A) Fluorescence photographs (for the GFP- or Rhodamine-selective channels) of representative cells taken 2 h after the cytochrome c injections. Arrows point to the GFP-positive cells that were injected with cytochrome c and rhodamine. (B) Quantitation of cell survival (2 h after the injections) for the various conditions. Data shown are the mean ± SEM of three independent experiments.

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Fluorescence, Injection, Quantitation Assay

    Apaf-1 but not caspase-9 levels are markedly reduced in cardiomyocytes in comparison to fibroblasts. (A) Western blots showing levels of Apaf-1 and caspase-9 proteins (and LDH and Troponin I as controls) in cultures of rat dermal fibroblasts and cardiomyocytes. (B) Quantitation of the data in which Apaf-1 and caspase-9 protein levels detected in cardiomyocyte cultures are expressed as a percentage of the levels (normalized to LDH) seen in fibroblast cultures. Data shown are the mean ± SEM of three independent experiments.
    Figure Legend Snippet: Apaf-1 but not caspase-9 levels are markedly reduced in cardiomyocytes in comparison to fibroblasts. (A) Western blots showing levels of Apaf-1 and caspase-9 proteins (and LDH and Troponin I as controls) in cultures of rat dermal fibroblasts and cardiomyocytes. (B) Quantitation of the data in which Apaf-1 and caspase-9 protein levels detected in cardiomyocyte cultures are expressed as a percentage of the levels (normalized to LDH) seen in fibroblast cultures. Data shown are the mean ± SEM of three independent experiments.

    Techniques Used: Western Blot, Quantitation Assay

    XIAP-deficient cardiomyocytes die with injection of cytochrome c . Cardiomyocytes isolated from XIAP-deficient (−/−) or wild-type (+/+) littermate mice were microinjected with 25 μg/μl of bovine cytochrome c . As a control, XIAP-deficient cardiomyocytes were also injected with 25 μg/μl of yeast cytochrome c . (A) Phase-contrast and fluorescence photographs of representative cells. Arrows mark the injected cells, identified by coinjection of rhodamine dextran along with cytochrome c . (B) Quantitation of cell survival at multiple times after the injections. Data shown are the mean ± SEM of three independent experiments.
    Figure Legend Snippet: XIAP-deficient cardiomyocytes die with injection of cytochrome c . Cardiomyocytes isolated from XIAP-deficient (−/−) or wild-type (+/+) littermate mice were microinjected with 25 μg/μl of bovine cytochrome c . As a control, XIAP-deficient cardiomyocytes were also injected with 25 μg/μl of yeast cytochrome c . (A) Phase-contrast and fluorescence photographs of representative cells. Arrows mark the injected cells, identified by coinjection of rhodamine dextran along with cytochrome c . (B) Quantitation of cell survival at multiple times after the injections. Data shown are the mean ± SEM of three independent experiments.

    Techniques Used: Injection, Isolation, Mouse Assay, Fluorescence, Quantitation Assay

    Cytosolic microinjection of cytochrome c induces death in fibroblasts but not cardiomyocytes. Neonatal rat cardiomyocytes and dermal fibroblasts were microinjected with 25 μg/μl of bovine or yeast cytochrome c , and cell survival (using morphological criteria) was assessed at multiple times after the injections. (A) Phase-contrast and fluorescence photographs of the cells 3 h after the injections of cytochrome c . The injected cells (arrows) were identified by the presence of rhodamine dextran coinjected with the cytochrome c . (B) Quantitation of cell survival. Data shown are the mean ± SEM of three independent experiments.
    Figure Legend Snippet: Cytosolic microinjection of cytochrome c induces death in fibroblasts but not cardiomyocytes. Neonatal rat cardiomyocytes and dermal fibroblasts were microinjected with 25 μg/μl of bovine or yeast cytochrome c , and cell survival (using morphological criteria) was assessed at multiple times after the injections. (A) Phase-contrast and fluorescence photographs of the cells 3 h after the injections of cytochrome c . The injected cells (arrows) were identified by the presence of rhodamine dextran coinjected with the cytochrome c . (B) Quantitation of cell survival. Data shown are the mean ± SEM of three independent experiments.

    Techniques Used: Fluorescence, Injection, Quantitation Assay

    Cardiomyocyte resistance to cytochrome c can be overcome with the exogenous addition of the IAP inhibitor Smac. (A) Western blots showing that the rat cardiomyocyte cultures express the core apoptotic components (Apaf-1, caspase-9, and caspase-3) and various IAPs (XIAP, cIAP-1, and cIAP-2). LDH and Troponin I are shown as controls. (B) Rat cardiomyocytes were microinjected with 25 μg/μl of bovine cytochrome c , 1 μg/μl of wild-type mature Smac, or both, and cell survival was assessed at multiple times after the injections. (C) Rat cardiomyocytes were injected with 25 μg/μl of bovine cytochrome c and 1 μg/μl of either mature wild-type AVPI-Smac or mature mutant MVPI-Smac in the presence or absence of 50 μM of the pan caspase inhibitor zVAD-FMK. Cell survival was assessed at multiple times after the injections. Data shown are the mean ± SEM of three independent experiments.
    Figure Legend Snippet: Cardiomyocyte resistance to cytochrome c can be overcome with the exogenous addition of the IAP inhibitor Smac. (A) Western blots showing that the rat cardiomyocyte cultures express the core apoptotic components (Apaf-1, caspase-9, and caspase-3) and various IAPs (XIAP, cIAP-1, and cIAP-2). LDH and Troponin I are shown as controls. (B) Rat cardiomyocytes were microinjected with 25 μg/μl of bovine cytochrome c , 1 μg/μl of wild-type mature Smac, or both, and cell survival was assessed at multiple times after the injections. (C) Rat cardiomyocytes were injected with 25 μg/μl of bovine cytochrome c and 1 μg/μl of either mature wild-type AVPI-Smac or mature mutant MVPI-Smac in the presence or absence of 50 μM of the pan caspase inhibitor zVAD-FMK. Cell survival was assessed at multiple times after the injections. Data shown are the mean ± SEM of three independent experiments.

    Techniques Used: Western Blot, Injection, Mutagenesis

    38) Product Images from "Skeletal Muscle Differentiation Evokes Endogenous XIAP to Restrict the Apoptotic Pathway"

    Article Title: Skeletal Muscle Differentiation Evokes Endogenous XIAP to Restrict the Apoptotic Pathway

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0005097

    tBid induced apoptosis in cardiomyocytes does not require endogenous Smac. (A) Rat cardiomyocytes were transfected with tBid-GFP or GFP alone. The percentage of transfected cells expressing active caspase 3 was determined 6 hours after transfection by immunohistochemistry. (B) Photographs of rat cardiomyocytes 6 hours after transfection with tBid-GFP in the presence or absence of the caspase inhibitor z-VAD-fmk (zVAD). (C) Wild type (wt) or Smac-deficient (Smac-/-) cardiomyocytes were infected with a tBid-GFP adenovirus in the presence or absence of z-VAD-fmk (zVAD). Cell survival was determined by morphology over a 12 hour period using time-lapse microscopy. (D) Protein levels of Smac were examined by Western blot of whole cell lysates from Smac-deficient dermal fibroblasts (Smac-/- DF), myotubes, cardiomyocytes and sympathetic neurons (neurons). Tubulin serves as a loading control. Densitometry of Smac protein levels are normalized to tubulin levels of the representative blot.
    Figure Legend Snippet: tBid induced apoptosis in cardiomyocytes does not require endogenous Smac. (A) Rat cardiomyocytes were transfected with tBid-GFP or GFP alone. The percentage of transfected cells expressing active caspase 3 was determined 6 hours after transfection by immunohistochemistry. (B) Photographs of rat cardiomyocytes 6 hours after transfection with tBid-GFP in the presence or absence of the caspase inhibitor z-VAD-fmk (zVAD). (C) Wild type (wt) or Smac-deficient (Smac-/-) cardiomyocytes were infected with a tBid-GFP adenovirus in the presence or absence of z-VAD-fmk (zVAD). Cell survival was determined by morphology over a 12 hour period using time-lapse microscopy. (D) Protein levels of Smac were examined by Western blot of whole cell lysates from Smac-deficient dermal fibroblasts (Smac-/- DF), myotubes, cardiomyocytes and sympathetic neurons (neurons). Tubulin serves as a loading control. Densitometry of Smac protein levels are normalized to tubulin levels of the representative blot.

    Techniques Used: Transfection, Expressing, Immunohistochemistry, Infection, Time-lapse Microscopy, Western Blot

    39) Product Images from "DNA single-strand break-induced DNA damage response causes heart failure"

    Article Title: DNA single-strand break-induced DNA damage response causes heart failure

    Journal: Nature Communications

    doi: 10.1038/ncomms15104

    Generation of an in vitro model of cardiomyocytes with SSB accumulation. ( a ) Neonatal rat cardiomyocytes (NRCMs) were treated with MMS at the indicated concentration for 10 min and the DNA damage was analysed by comet assay (Alkaline comet: n =42, 37, 45, 33, 34; Neutral comet: n =40, 35, 35, 37, 29 at each concentration, respectively). Statistical significance was determined by Steel–Dwass test. ## P
    Figure Legend Snippet: Generation of an in vitro model of cardiomyocytes with SSB accumulation. ( a ) Neonatal rat cardiomyocytes (NRCMs) were treated with MMS at the indicated concentration for 10 min and the DNA damage was analysed by comet assay (Alkaline comet: n =42, 37, 45, 33, 34; Neutral comet: n =40, 35, 35, 37, 29 at each concentration, respectively). Statistical significance was determined by Steel–Dwass test. ## P

    Techniques Used: In Vitro, Concentration Assay, Single Cell Gel Electrophoresis

    Possible roles of SSB accumulation in pathogenesis of heart failure. Accumulation of DNA SSB in cardiomyocytes induces persistent activation of DDR and subsequent activation of NF-κB pathway, resulting in increased expressions of inflammatory cytokines. These mechanisms may contribute, at least in part, to increased cardiac inflammation and the progression of pressure overload-induced heart failure.
    Figure Legend Snippet: Possible roles of SSB accumulation in pathogenesis of heart failure. Accumulation of DNA SSB in cardiomyocytes induces persistent activation of DDR and subsequent activation of NF-κB pathway, resulting in increased expressions of inflammatory cytokines. These mechanisms may contribute, at least in part, to increased cardiac inflammation and the progression of pressure overload-induced heart failure.

    Techniques Used: Activation Assay

    Xrcc1 deficiency exacerbates cardiac inflammation after pressure overload. ( a , b ) Activation of DDR in Sham- or TAC-operated Xrcc1 f/f and Xrcc1 αMHC-Cre mice was assessed by immunostaining for phosphorylated H2AX ( a , γH2AX, green). Immunostaining for alpha-actinin (red) was used to label cardiomyocytes. Arrowheads indicate γH2AX-positive cardiomyocytes and arrows indicate γH2AX-positive non-cardiomyocytes. Scale bar, 50 μm. The number of γH2AX-positive cardiomyocytes was counted ( b , n =4 each). Statistical significance was determined by one-way analysis of variance (ANOVA) followed by the Tukey–Kramer HSD test. ** P
    Figure Legend Snippet: Xrcc1 deficiency exacerbates cardiac inflammation after pressure overload. ( a , b ) Activation of DDR in Sham- or TAC-operated Xrcc1 f/f and Xrcc1 αMHC-Cre mice was assessed by immunostaining for phosphorylated H2AX ( a , γH2AX, green). Immunostaining for alpha-actinin (red) was used to label cardiomyocytes. Arrowheads indicate γH2AX-positive cardiomyocytes and arrows indicate γH2AX-positive non-cardiomyocytes. Scale bar, 50 μm. The number of γH2AX-positive cardiomyocytes was counted ( b , n =4 each). Statistical significance was determined by one-way analysis of variance (ANOVA) followed by the Tukey–Kramer HSD test. ** P

    Techniques Used: Activation Assay, Mouse Assay, Immunostaining

    Accumulation of DNA SSB in the failing heart. ( a , b ) Cardiomyocytes were isolated from the TAC-operated heart at the indicated time points. The type of DNA damage in cardiomyocytes was assessed by comet assay. Representative images ( a ) and quantitative analyses are shown ( b , Alkaline comet: n =28, 45, 48; Neutral comet: n =38, 56, 44 at each time point, respectively, biological replicates=3). ( c , d ) Fragmented DNA and DSB were labelled with ISOL staining ( c , green). Wheat germ agglutinin (WGA, red) was used to visualize cardiomyocytes. DNase-treated section (DNase I, 10 Kunitz units ml −1 ) was used as a positive control. Arrowheads indicate ISOL-positive cardiomyocytes and arrows indicate ISOL-positive non-cardiomyocytes. White scale bar, 50 μm; yellow scale bar, 20 μm. The number of ISOL-positive cardiomyocytes was counted ( d , n =4 each). ( e , f ) Heart tissue sections were immunostained for NBS1 ( e , NBS1, green). Immunostaining for alpha-actinin (red) was used to label cardiomyocytes. Scale bar, 50 μm. The number of NBS1-positive cardiomyocytes was counted ( f , n =4 each). ( g , h ) Heart tissue sections were immunostained for poly-ADP ribose ( g , PAR, green) and the number of PAR-positive cardiomyocytes was counted ( h , n =4, 4, 5 at each time point, respectively). Arrowheads indicate PAR-positive cardiomyocytes and arrows indicate PAR-positive non-cardiomyocytes. White scale bar, 50 μm; yellow scale bar, 20 μm. ( i ) Expression levels of SSB repair enzymes were analysed by real-time PCR ( n =4, 6, 8 at each time point, respectively, technical duplicates). ( j , k ) Heart tissue sections were stained with dihydroethidium ( i , DHE, 10 μΜ) and mean fluorescence intensity relative to Sham-operated mice was measured ( k , n =4, 5, 5 at each time point, respectively). Scale bar, 50 μm. ( l ) The level of H 2 O 2 in the TAC-operated heart was measured using Amplex Red assay ( n =9, 5, 6 at each time point, respectively). Statistical significance was determined by Steel-Dwass test for ( b ) and by one-way analysis of variance followed by the Tukey–Kramer HSD test for ( d , f , h , i , j ) * P
    Figure Legend Snippet: Accumulation of DNA SSB in the failing heart. ( a , b ) Cardiomyocytes were isolated from the TAC-operated heart at the indicated time points. The type of DNA damage in cardiomyocytes was assessed by comet assay. Representative images ( a ) and quantitative analyses are shown ( b , Alkaline comet: n =28, 45, 48; Neutral comet: n =38, 56, 44 at each time point, respectively, biological replicates=3). ( c , d ) Fragmented DNA and DSB were labelled with ISOL staining ( c , green). Wheat germ agglutinin (WGA, red) was used to visualize cardiomyocytes. DNase-treated section (DNase I, 10 Kunitz units ml −1 ) was used as a positive control. Arrowheads indicate ISOL-positive cardiomyocytes and arrows indicate ISOL-positive non-cardiomyocytes. White scale bar, 50 μm; yellow scale bar, 20 μm. The number of ISOL-positive cardiomyocytes was counted ( d , n =4 each). ( e , f ) Heart tissue sections were immunostained for NBS1 ( e , NBS1, green). Immunostaining for alpha-actinin (red) was used to label cardiomyocytes. Scale bar, 50 μm. The number of NBS1-positive cardiomyocytes was counted ( f , n =4 each). ( g , h ) Heart tissue sections were immunostained for poly-ADP ribose ( g , PAR, green) and the number of PAR-positive cardiomyocytes was counted ( h , n =4, 4, 5 at each time point, respectively). Arrowheads indicate PAR-positive cardiomyocytes and arrows indicate PAR-positive non-cardiomyocytes. White scale bar, 50 μm; yellow scale bar, 20 μm. ( i ) Expression levels of SSB repair enzymes were analysed by real-time PCR ( n =4, 6, 8 at each time point, respectively, technical duplicates). ( j , k ) Heart tissue sections were stained with dihydroethidium ( i , DHE, 10 μΜ) and mean fluorescence intensity relative to Sham-operated mice was measured ( k , n =4, 5, 5 at each time point, respectively). Scale bar, 50 μm. ( l ) The level of H 2 O 2 in the TAC-operated heart was measured using Amplex Red assay ( n =9, 5, 6 at each time point, respectively). Statistical significance was determined by Steel-Dwass test for ( b ) and by one-way analysis of variance followed by the Tukey–Kramer HSD test for ( d , f , h , i , j ) * P

    Techniques Used: Isolation, Single Cell Gel Electrophoresis, Staining, Whole Genome Amplification, Positive Control, Immunostaining, Expressing, Real-time Polymerase Chain Reaction, Fluorescence, Mouse Assay, Amplex Red Assay

    ATM gene deletion rescues the cardiac phenotypes of Xrcc1 deficient mice. ( a , b ) Macroscopic and echocardiographic images ( a ) and cardiac function ( b ) of TAC-operated Xrcc1 f/f , Xrcc1 αMHC-Cre and Xrcc1 αMHC-Cre ; Atm +/− mice ( Xrcc1 f/f mice: n =83, 21, 46, 11, 27; Xrcc1 αMHC-Cre mice: n =88, 28, 60, 13, 16; Xrcc1 αMHC-Cre ; Atm +/− mice: n =28, 22, 22, 7, 7 at each time point, respectively). Scale bar, 2 mm. ( c ) Heart, lung, and body weight of TAC-operated Xrcc1 f/f , Xrcc1 αMHC-Cre and Xrcc1 αMHC-Cre ; Atm +/− mice were weighed 8 weeks after the surgery ( n =8, 5, 6 for each genotype, respectively). ( d ) Survival curves of TAC-operated Xrcc1 f/f , Xrcc1 αMHC-Cre and Xrcc1 αMHC-Cre ; Atm +/− mice ( n =49, 62, 23, respectively). ( e – k ) TAC-operated Xrcc1 f/f , Xrcc1 αMHC-Cre and Xrcc1 αMHC-Cre ; Atm +/− mice were analysed 4 weeks after the surgery. The type of DNA damage in cardiomyocytes was assessed by comet assay ( e , Alkaline comet: n =50, 76, 77; Neutral comet: n =53, 56, 42, respectively). Activation of DDR was assessed by immunostaining for phosphorylated H2AX ( f , γH2AX, green, arrowheads). Arrowheads indicate γH2AX-positive cardiomyocytes and arrows indicate γH2AX-positive non-cardiomyocytes. Scale bar, 50 μm. The number of γH2AX-positive cardiomyocytes was counted ( g , n =4 each). Expression levels of inflammatory cytokines in the isolated cardiomyocytes were assessed by real-time PCR ( h , n =10, 16, 12 for each genotype, respectively, technical duplicates). ChIP–qPCR analysis of binding of NF-κB to the Vcam1 promoter region. Data is presented as fold enrichment relative to TAC-operated Xrcc1 f/f mice ( i , n =4, 5, 5, respectively). Heart tissues were immunostained for CD45 or CD68 ( j , green, arrowheads). Arrowheads indicate CD45- or CD68-positive cells. Scale bar, 50 μm. The number of CD45- and CD68-positive cells was counted ( k , n =4 each). Statistical significance was determined by one-way analysis of variance followed by the Tukey–Kramer HSD test for ( b ) (at each time point), ( c , g , h , i , k ), by Wilcoxon test for d and by Steel–Dwass test for e , # P
    Figure Legend Snippet: ATM gene deletion rescues the cardiac phenotypes of Xrcc1 deficient mice. ( a , b ) Macroscopic and echocardiographic images ( a ) and cardiac function ( b ) of TAC-operated Xrcc1 f/f , Xrcc1 αMHC-Cre and Xrcc1 αMHC-Cre ; Atm +/− mice ( Xrcc1 f/f mice: n =83, 21, 46, 11, 27; Xrcc1 αMHC-Cre mice: n =88, 28, 60, 13, 16; Xrcc1 αMHC-Cre ; Atm +/− mice: n =28, 22, 22, 7, 7 at each time point, respectively). Scale bar, 2 mm. ( c ) Heart, lung, and body weight of TAC-operated Xrcc1 f/f , Xrcc1 αMHC-Cre and Xrcc1 αMHC-Cre ; Atm +/− mice were weighed 8 weeks after the surgery ( n =8, 5, 6 for each genotype, respectively). ( d ) Survival curves of TAC-operated Xrcc1 f/f , Xrcc1 αMHC-Cre and Xrcc1 αMHC-Cre ; Atm +/− mice ( n =49, 62, 23, respectively). ( e – k ) TAC-operated Xrcc1 f/f , Xrcc1 αMHC-Cre and Xrcc1 αMHC-Cre ; Atm +/− mice were analysed 4 weeks after the surgery. The type of DNA damage in cardiomyocytes was assessed by comet assay ( e , Alkaline comet: n =50, 76, 77; Neutral comet: n =53, 56, 42, respectively). Activation of DDR was assessed by immunostaining for phosphorylated H2AX ( f , γH2AX, green, arrowheads). Arrowheads indicate γH2AX-positive cardiomyocytes and arrows indicate γH2AX-positive non-cardiomyocytes. Scale bar, 50 μm. The number of γH2AX-positive cardiomyocytes was counted ( g , n =4 each). Expression levels of inflammatory cytokines in the isolated cardiomyocytes were assessed by real-time PCR ( h , n =10, 16, 12 for each genotype, respectively, technical duplicates). ChIP–qPCR analysis of binding of NF-κB to the Vcam1 promoter region. Data is presented as fold enrichment relative to TAC-operated Xrcc1 f/f mice ( i , n =4, 5, 5, respectively). Heart tissues were immunostained for CD45 or CD68 ( j , green, arrowheads). Arrowheads indicate CD45- or CD68-positive cells. Scale bar, 50 μm. The number of CD45- and CD68-positive cells was counted ( k , n =4 each). Statistical significance was determined by one-way analysis of variance followed by the Tukey–Kramer HSD test for ( b ) (at each time point), ( c , g , h , i , k ), by Wilcoxon test for d and by Steel–Dwass test for e , # P

    Techniques Used: Mouse Assay, Single Cell Gel Electrophoresis, Activation Assay, Immunostaining, Expressing, Isolation, Real-time Polymerase Chain Reaction, Chromatin Immunoprecipitation, Binding Assay

    40) Product Images from "Heat‐shock transcription factor 1 is critically involved in the ischaemia‐induced cardiac hypertrophy via JAK2/ STAT3 pathway, et al. Heat‐shock transcription factor 1 is critically involved in the ischaemia‐induced cardiac hypertrophy via JAK2/STAT3 pathway"

    Article Title: Heat‐shock transcription factor 1 is critically involved in the ischaemia‐induced cardiac hypertrophy via JAK2/ STAT3 pathway, et al. Heat‐shock transcription factor 1 is critically involved in the ischaemia‐induced cardiac hypertrophy via JAK2/STAT3 pathway

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.13713

    Deficiency of HSF 1 deteriorated hypoxia‐induced myocardial hypertrophy in vitro. A,. Representative Western blots of total and phosphorylation level of HSF 1 in neonatal cardiomyocytes ( NRCM s) after transduced with indicated Si RNA and treated with hypoxia. N = 5 independent experiments. B,. Quantitative results of total and phosphorylation level of HSF 1in NRCM s. C and D,. The representative images and quantification data of cardiomyocyte surface area ( CSA ) of NRCM s stained by α‐myosin heavy chain (α‐ MHC ). Scale bar: 20 μm. E, mRNA levels of hypertrophic markers, atrial natriuretic peptide ( ANP ), B‐type natriuretic peptide ( BNP ) in NRCM s measured by real‐time PCR . GAPDH was served as internal control. F and G,. The TUNEL staining and quantified data of NRCM s. Scale bar: 20 μm. N = 5 independent experiments. NO , normoxia. HO , hypoxia. All data are expressed as mean ± SEM * P
    Figure Legend Snippet: Deficiency of HSF 1 deteriorated hypoxia‐induced myocardial hypertrophy in vitro. A,. Representative Western blots of total and phosphorylation level of HSF 1 in neonatal cardiomyocytes ( NRCM s) after transduced with indicated Si RNA and treated with hypoxia. N = 5 independent experiments. B,. Quantitative results of total and phosphorylation level of HSF 1in NRCM s. C and D,. The representative images and quantification data of cardiomyocyte surface area ( CSA ) of NRCM s stained by α‐myosin heavy chain (α‐ MHC ). Scale bar: 20 μm. E, mRNA levels of hypertrophic markers, atrial natriuretic peptide ( ANP ), B‐type natriuretic peptide ( BNP ) in NRCM s measured by real‐time PCR . GAPDH was served as internal control. F and G,. The TUNEL staining and quantified data of NRCM s. Scale bar: 20 μm. N = 5 independent experiments. NO , normoxia. HO , hypoxia. All data are expressed as mean ± SEM * P

    Techniques Used: In Vitro, Western Blot, Transduction, Staining, Aqueous Normal-phase Chromatography, Real-time Polymerase Chain Reaction, TUNEL Assay

    Enhance of HSF 1 attenuated cardiac hypertrophy induced by MI in mice. A, Heart weight/body weight ratio ( HW / BW ), (B) left ventricular weight/tibia length ratio ( LVW / TL ) and (C) lung weight/tibia length ratio ( LW / TL ) of mice. D, Representative images of histological sections of the mouse LV s were stained with haematoxylin‐eosin (H E) 1 wk after MI or sham surgery. Scale bar: 50 μm. E, Quantitative results of the cross‐sectional area ( CSA ) of mouse cardiomyocytes quantified using an image analysis system. F, Real‐time PCR analysis of ANP and BNP in mouse border zone area of LV s 1 wk after MI . (G) The infarction size of the mouse hearts. H and I, The TUNEL staining and quantified data of mouse hearts. Scale bar: 50μm. GAPDH was served as internal control. N = 7 per experimental group. All data are expressed as mean ± SEM * P
    Figure Legend Snippet: Enhance of HSF 1 attenuated cardiac hypertrophy induced by MI in mice. A, Heart weight/body weight ratio ( HW / BW ), (B) left ventricular weight/tibia length ratio ( LVW / TL ) and (C) lung weight/tibia length ratio ( LW / TL ) of mice. D, Representative images of histological sections of the mouse LV s were stained with haematoxylin‐eosin (H E) 1 wk after MI or sham surgery. Scale bar: 50 μm. E, Quantitative results of the cross‐sectional area ( CSA ) of mouse cardiomyocytes quantified using an image analysis system. F, Real‐time PCR analysis of ANP and BNP in mouse border zone area of LV s 1 wk after MI . (G) The infarction size of the mouse hearts. H and I, The TUNEL staining and quantified data of mouse hearts. Scale bar: 50μm. GAPDH was served as internal control. N = 7 per experimental group. All data are expressed as mean ± SEM * P

    Techniques Used: Mouse Assay, Staining, Real-time Polymerase Chain Reaction, Aqueous Normal-phase Chromatography, TUNEL Assay

    Deficiency of HSF 1 aggravated cardiac hypertrophy induced by MI in mice. A, Heart weight/body weight ratio ( HW / BW ) (B) left ventricular weight/tibia length ratio ( LVW / TL ) and (C) lung weight/tibia length ratio ( LW / TL ) of mice. D, Representative images of histological sections of the mouse LV s were stained with haematoxylin‐eosin (H E) 1 wk after MI or sham surgery. Scale bar: 50 μm. E, Quantitative results of the cross‐sectional area ( CSA ) of mouse cardiomyocytes quantified using an image analysis system. F,. Real‐time PCR analysis of ANP and BNP in mouse border zone area of LV s 1 wk after MI . G, the infarction size of the mouse hearts. H and I, The TUNEL staining and quantified data of mouse hearts. Scale bar: 20 μm. GAPDH was served as internal control. N = 7‐8 per experimental group. All data are expressed as mean ± SEM * P
    Figure Legend Snippet: Deficiency of HSF 1 aggravated cardiac hypertrophy induced by MI in mice. A, Heart weight/body weight ratio ( HW / BW ) (B) left ventricular weight/tibia length ratio ( LVW / TL ) and (C) lung weight/tibia length ratio ( LW / TL ) of mice. D, Representative images of histological sections of the mouse LV s were stained with haematoxylin‐eosin (H E) 1 wk after MI or sham surgery. Scale bar: 50 μm. E, Quantitative results of the cross‐sectional area ( CSA ) of mouse cardiomyocytes quantified using an image analysis system. F,. Real‐time PCR analysis of ANP and BNP in mouse border zone area of LV s 1 wk after MI . G, the infarction size of the mouse hearts. H and I, The TUNEL staining and quantified data of mouse hearts. Scale bar: 20 μm. GAPDH was served as internal control. N = 7‐8 per experimental group. All data are expressed as mean ± SEM * P

    Techniques Used: Mouse Assay, Staining, Real-time Polymerase Chain Reaction, Aqueous Normal-phase Chromatography, TUNEL Assay

    Related Articles

    Patch Clamp:

    Article Title: Stilbene derivative as a photosensitive compound to control the excitability of neonatal rat cardiomyocytes
    Article Snippet: .. Preparation of neonatal rat ventricular cardiomyocyte (NRVM) for patch clamp for optical mapping Hearts were isolated from neonatal 1–2-day-old Wistar rats using a two-day protocol of Neonatal Cardiomyocyte Isolation System of Worthington Biochemical Corporation [ ] with some modifications. ..

    Isolation:

    Article Title: Three-Dimensional Culture Alters Primary Cardiac Cell Phenotype
    Article Snippet: .. NRCVCs were isolated using Neonatal Cardiomyocyte Isolation System kits (Worthington Biochemical, Lakewood, NJ) as in previous studies by our group. .. Viable cells were enumerated by Trypan-blue exclusion (Invitrogen, Carlsbad, CA).

    Article Title: Single Cell Transcriptomics Reconstructs Fate Conversion from Fibroblast to Cardiomyocyte
    Article Snippet: .. Neonatal CMs were isolated using the neonatal cardiomyocytes isolation system (Worthington Biochemical Corporation) except that all enzymes were used at a ¼ of the recommended concentration to increase cell viability. .. After a 1.5 hour pre-plating on uncoated surface to remove attached nonmyocytes, the unattached CMs were collected in TRIzol ( > 80% viability by Trypan blue staining).

    Article Title: Control of histone H3 phosphorylation by CaMKIIδ in response to haemodynamic cardiac stress
    Article Snippet: .. Primary neonatal rat cardiomyocytes isolated using the Worthington Neonatal CardioMyocytes System (Worthington, USA) were co-transfected with a GFP-tagged plasmid and siControl (siCt) or siRNA against 14–3–3 (si14–3–3) from Dharmacon (Thermo Scientific, USA) using lipofectamine (Invitrogen). .. 24 h post-transfection, total cell lysates were prepared and analysed by western blotting or immunofluorescence.

    Article Title: Human Myocardial Pericytes: Multipotent Mesodermal Precursors Exhibiting Cardiac Specificity
    Article Snippet: .. Rat neonatal cardiomyocytes were isolated from anaesthetized 3-day-old rat pups using the Worthington rat neonatal cardiomyocyte isolation system per the manufacturer’s protocol (Worthington Biochemical Corp). .. Pericyte/cardiomyocyte co-culture protocol was adopted and modified from previous reports ( , ).

    Article Title: Muscle ring finger protein-1 inhibits PKC? activation and prevents cardiomyocyte hypertrophy
    Article Snippet: .. NRVM were isolated using the neonatal cardiomyocyte isolation kit (Worthington) and were plated on laminin. ..

    Article Title: An siRNA Screen Identifies the U2 snRNP Spliceosome as a Host Restriction Factor for Recombinant Adeno-associated Viruses
    Article Snippet: .. Primary cardiomyocytes were isolated from newborn Dahl salt-sensitive rats using the Neonatal Cardiomyocytes Isolation System (Worthington, Lakewood, NJ) according to the manufacturer’s instruction. ..

    Article Title: Pivotal role of cardiac lineage protein-1 (CLP-1) in transcriptional elongation factor P-TEFb complex formation in cardiac hypertrophy
    Article Snippet: .. Cardiac cells of 2–4-day-old WKY rats were isolated using a neonatal cardiomyocyte isolation system (Worthington Biochemical Corp, Lakewood, New Jersey, USA). .. After pre-plating to reduce contaminating non-muscle cells, cardiomyocytes were plated on Bioflex collagen 6-well plates (Flexcell, McKeesport, Pennsylvania, USA) and cultured in DMEM/F-12 (1:1) media supplemented with 10% fetal bovine serum for approximately 1.5 days prior to switching to serum-free medium.

    Article Title: Stilbene derivative as a photosensitive compound to control the excitability of neonatal rat cardiomyocytes
    Article Snippet: .. Preparation of neonatal rat ventricular cardiomyocyte (NRVM) for patch clamp for optical mapping Hearts were isolated from neonatal 1–2-day-old Wistar rats using a two-day protocol of Neonatal Cardiomyocyte Isolation System of Worthington Biochemical Corporation [ ] with some modifications. ..

    Plasmid Preparation:

    Article Title: Control of histone H3 phosphorylation by CaMKIIδ in response to haemodynamic cardiac stress
    Article Snippet: .. Primary neonatal rat cardiomyocytes isolated using the Worthington Neonatal CardioMyocytes System (Worthington, USA) were co-transfected with a GFP-tagged plasmid and siControl (siCt) or siRNA against 14–3–3 (si14–3–3) from Dharmacon (Thermo Scientific, USA) using lipofectamine (Invitrogen). .. 24 h post-transfection, total cell lysates were prepared and analysed by western blotting or immunofluorescence.

    Concentration Assay:

    Article Title: Single Cell Transcriptomics Reconstructs Fate Conversion from Fibroblast to Cardiomyocyte
    Article Snippet: .. Neonatal CMs were isolated using the neonatal cardiomyocytes isolation system (Worthington Biochemical Corporation) except that all enzymes were used at a ¼ of the recommended concentration to increase cell viability. .. After a 1.5 hour pre-plating on uncoated surface to remove attached nonmyocytes, the unattached CMs were collected in TRIzol ( > 80% viability by Trypan blue staining).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93
    Worthington Biochemical horizontal run up velocity
    Horizontal Run Up Velocity, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/horizontal run up velocity/product/Worthington Biochemical
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    horizontal run up velocity - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    Image Search Results