hcn4  (Alomone Labs)


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    Name:
    Anti HCN4 Antibody
    Description:
    Anti HCN4 Antibody APC 052 is a highly specific antibody directed against an epitope of the human protein The antibody can be used in western blot immunoprecipitation immunocytochemistry and immunohistochemistry applications It has been designed to recognize HCN4 from human rat and mouse samples
    Catalog Number:
    APC-052
    Price:
    397.0
    Category:
    Primary Antibody
    Applications:
    Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western Blot
    Purity:
    The serum was depleted of anti-GST antibodies by affinity chromatography on immobilized GST and then the IgG fraction was purified on immobilized antigen.
    Immunogen:
    GST fusion protein
    Size:
    25 mcl
    Antibody Type:
    Polyclonal Primary Antibodies
    Format:
    Lyophilized Powder
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
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    Structured Review

    Alomone Labs hcn4
    Anti HCN4 Antibody
    Anti HCN4 Antibody APC 052 is a highly specific antibody directed against an epitope of the human protein The antibody can be used in western blot immunoprecipitation immunocytochemistry and immunohistochemistry applications It has been designed to recognize HCN4 from human rat and mouse samples
    https://www.bioz.com/result/hcn4/product/Alomone Labs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hcn4 - by Bioz Stars, 2021-09
    95/100 stars

    Images

    1) Product Images from "MyoR Modulates Cardiac Conduction by Repressing Gata4"

    Article Title: MyoR Modulates Cardiac Conduction by Repressing Gata4

    Journal: Molecular and Cellular Biology

    doi: 10.1128/MCB.00860-14

    Transcriptional profiling of Cx30.2-lacZ + cells identifies MyoR as an AVC-enriched transcript. (A) Strategy used for purifying E12.5 Cx30.2-lacZ + AVC cells by flow cytometry. (B) Partial list of genes found to be enriched in microarray analysis. Fold enrichment refers to transcript levels in Cx30.2-lacZ + cells relative to lacZ -null cells. Note that MyoR transcripts were 7-fold more abundant in Cx30.2-lacZ + cells within the E12.5 heart. (C) qRT-PCR analysis of microdissected E10.5 AVC tissue demonstrating enrichment of MyoR. Fold enrichment refers to transcript levels in dissected AVC tissue versus whole heart. (D) Developmental analysis of MyoR expression by qRT-PCR analysis. MyoR expression in the whole heart at a particular time point relative to E10.5 is shown. MyoR transcripts were found to peak during late gestation (E16.5). (E) qRT-PCR analysis of MyoR expression level in Cx30.2-EGFP + AVC cells isolated at E16.5. MyoR expression is shown relative to transcript levels in EGFP − AVC cells. (F) Western blot analysis of AVC and ventricular nuclear extracts dissected from E16.5 embryos (top) with the indicated antibodies. Band quantitation and normalization demonstrated ∼14-fold enrichment of MyoR in the E16.5 AVC relative to ventricular tissue. (G) Immunostaining of consecutive sections through an E16.5 Cx30.2-EGFP heart for GFP, Gata4, Hcn4, and MyoR, demonstrating localization of Cx30.2, Gata4, and MyoR to the Hcn4 + AVN (dashed outline). All sections were counterstained with DAPI to stain cell nuclei. Scale bar, 20 μm. LV, left ventricle; RV, right ventricle; FDG, fluorescein digalactopyranoside; AVC, atrioventricular canal; AVN, atrioventricular node.
    Figure Legend Snippet: Transcriptional profiling of Cx30.2-lacZ + cells identifies MyoR as an AVC-enriched transcript. (A) Strategy used for purifying E12.5 Cx30.2-lacZ + AVC cells by flow cytometry. (B) Partial list of genes found to be enriched in microarray analysis. Fold enrichment refers to transcript levels in Cx30.2-lacZ + cells relative to lacZ -null cells. Note that MyoR transcripts were 7-fold more abundant in Cx30.2-lacZ + cells within the E12.5 heart. (C) qRT-PCR analysis of microdissected E10.5 AVC tissue demonstrating enrichment of MyoR. Fold enrichment refers to transcript levels in dissected AVC tissue versus whole heart. (D) Developmental analysis of MyoR expression by qRT-PCR analysis. MyoR expression in the whole heart at a particular time point relative to E10.5 is shown. MyoR transcripts were found to peak during late gestation (E16.5). (E) qRT-PCR analysis of MyoR expression level in Cx30.2-EGFP + AVC cells isolated at E16.5. MyoR expression is shown relative to transcript levels in EGFP − AVC cells. (F) Western blot analysis of AVC and ventricular nuclear extracts dissected from E16.5 embryos (top) with the indicated antibodies. Band quantitation and normalization demonstrated ∼14-fold enrichment of MyoR in the E16.5 AVC relative to ventricular tissue. (G) Immunostaining of consecutive sections through an E16.5 Cx30.2-EGFP heart for GFP, Gata4, Hcn4, and MyoR, demonstrating localization of Cx30.2, Gata4, and MyoR to the Hcn4 + AVN (dashed outline). All sections were counterstained with DAPI to stain cell nuclei. Scale bar, 20 μm. LV, left ventricle; RV, right ventricle; FDG, fluorescein digalactopyranoside; AVC, atrioventricular canal; AVN, atrioventricular node.

    Techniques Used: Flow Cytometry, Cytometry, Microarray, Quantitative RT-PCR, Expressing, Isolation, Western Blot, Quantitation Assay, Immunostaining, Staining

    2) Product Images from "Spatiotemporal Regulation of an Hcn4 Enhancer Defines a Role for Mef2c and HDACs in Cardiac Electrical Patterning"

    Article Title: Spatiotemporal Regulation of an Hcn4 Enhancer Defines a Role for Mef2c and HDACs in Cardiac Electrical Patterning

    Journal: Developmental biology

    doi: 10.1016/j.ydbio.2012.10.017

    Identification of minimal enhancer regions within R2R3 that direct reporter activity in the Hcn4 expression domain
    Figure Legend Snippet: Identification of minimal enhancer regions within R2R3 that direct reporter activity in the Hcn4 expression domain

    Techniques Used: Activity Assay, Expressing

    HDAC Activity Regulates Hcn4 R2R3
    Figure Legend Snippet: HDAC Activity Regulates Hcn4 R2R3

    Techniques Used: Activity Assay

    Mef2C regulates R2R3-LacZ and Hcn4 expression in vivo
    Figure Legend Snippet: Mef2C regulates R2R3-LacZ and Hcn4 expression in vivo

    Techniques Used: Expressing, In Vivo

    Identification and in vivo testing of Hcn4 regulatory elements
    Figure Legend Snippet: Identification and in vivo testing of Hcn4 regulatory elements

    Techniques Used: In Vivo

    In vivo temporal reporter activity directed by the combined R2-R3 Hcn4 Enhancers
    Figure Legend Snippet: In vivo temporal reporter activity directed by the combined R2-R3 Hcn4 Enhancers

    Techniques Used: In Vivo, Activity Assay

    In vivo expression directed by the R2R3 Hcn4 enhancer in the post-natal AV bundle and its interface with the AV Node
    Figure Legend Snippet: In vivo expression directed by the R2R3 Hcn4 enhancer in the post-natal AV bundle and its interface with the AV Node

    Techniques Used: In Vivo, Expressing

    Mef2C regulates R2R3 Hcn4 enhancer activity via a conserved Mef2 Site
    Figure Legend Snippet: Mef2C regulates R2R3 Hcn4 enhancer activity via a conserved Mef2 Site

    Techniques Used: Activity Assay

    Expression pattern of Hcn4 mRNA by whole-mount in-situ hybridization
    Figure Legend Snippet: Expression pattern of Hcn4 mRNA by whole-mount in-situ hybridization

    Techniques Used: Expressing, In Situ Hybridization

    3) Product Images from "Programming and Isolation of Highly Pure Physiologically and Pharmacologically Functional Sinus-Nodal Bodies from Pluripotent Stem Cells"

    Article Title: Programming and Isolation of Highly Pure Physiologically and Pharmacologically Functional Sinus-Nodal Bodies from Pluripotent Stem Cells

    Journal: Stem Cell Reports

    doi: 10.1016/j.stemcr.2014.03.006

    Electrophysiological Parameters of Pacemaker-like Cardiomyocytes Obtained from iSABs (A) Distribution of pacemaker cells as shown by single-cell patch-clamp and funny-channel measurements: > 81% of pacemaker-like cells were achieved; 19% of these represented immature pacemaker cells and the rest were mature pacemaker cells (n = 65). The data represent seven independent experiments and are presented as means ± SD. See also Tables S1–S3 . (B) Patch-clamp protocol. Voltage was applied to elicit the hyperpolarization-activated current for I f recordings from isolated iSAB cells. (C) Example of I f current, recorded from an isolated iSAB-derived cell. (D and E) Current density (D) and time constant of activation (E) at –130 mV, demonstrating a robust I f expression with slow activation kinetics typical of the HCN4 channel subtype and mature SA nodal I f (n = 17). The data represent three independent experiments and are presented as means ± SD. (F) Reaction to β-adrenergic (isoprotenerol) and muscarinic (carbachol) stimulation leads to typical accelerated versus decelerated AP rates. iSAB-derived cells show a pronounced response to isoprotenerol, with beating rates reaching up to 560 bpm. iSAB/Iso: n = 9; iSAB/Carb: n = 5. The data represent seven independent experiments and are presented as means ± SD. See also Figures S1 and S2 .
    Figure Legend Snippet: Electrophysiological Parameters of Pacemaker-like Cardiomyocytes Obtained from iSABs (A) Distribution of pacemaker cells as shown by single-cell patch-clamp and funny-channel measurements: > 81% of pacemaker-like cells were achieved; 19% of these represented immature pacemaker cells and the rest were mature pacemaker cells (n = 65). The data represent seven independent experiments and are presented as means ± SD. See also Tables S1–S3 . (B) Patch-clamp protocol. Voltage was applied to elicit the hyperpolarization-activated current for I f recordings from isolated iSAB cells. (C) Example of I f current, recorded from an isolated iSAB-derived cell. (D and E) Current density (D) and time constant of activation (E) at –130 mV, demonstrating a robust I f expression with slow activation kinetics typical of the HCN4 channel subtype and mature SA nodal I f (n = 17). The data represent three independent experiments and are presented as means ± SD. (F) Reaction to β-adrenergic (isoprotenerol) and muscarinic (carbachol) stimulation leads to typical accelerated versus decelerated AP rates. iSAB-derived cells show a pronounced response to isoprotenerol, with beating rates reaching up to 560 bpm. iSAB/Iso: n = 9; iSAB/Carb: n = 5. The data represent seven independent experiments and are presented as means ± SD. See also Figures S1 and S2 .

    Techniques Used: Patch Clamp, Isolation, Derivative Assay, Activation Assay, Expressing

    4) Product Images from "Programming and Isolation of Highly Pure Physiologically and Pharmacologically Functional Sinus-Nodal Bodies from Pluripotent Stem Cells"

    Article Title: Programming and Isolation of Highly Pure Physiologically and Pharmacologically Functional Sinus-Nodal Bodies from Pluripotent Stem Cells

    Journal: Stem Cell Reports

    doi: 10.1016/j.stemcr.2014.03.006

    Electrophysiological Parameters of Pacemaker-like Cardiomyocytes Obtained from iSABs (A) Distribution of pacemaker cells as shown by single-cell patch-clamp and funny-channel measurements: > 81% of pacemaker-like cells were achieved; 19% of these represented immature pacemaker cells and the rest were mature pacemaker cells (n = 65). The data represent seven independent experiments and are presented as means ± SD. See also Tables S1–S3 . (B) Patch-clamp protocol. Voltage was applied to elicit the hyperpolarization-activated current for I f recordings from isolated iSAB cells. (C) Example of I f current, recorded from an isolated iSAB-derived cell. (D and E) Current density (D) and time constant of activation (E) at –130 mV, demonstrating a robust I f expression with slow activation kinetics typical of the HCN4 channel subtype and mature SA nodal I f (n = 17). The data represent three independent experiments and are presented as means ± SD. (F) Reaction to β-adrenergic (isoprotenerol) and muscarinic (carbachol) stimulation leads to typical accelerated versus decelerated AP rates. iSAB-derived cells show a pronounced response to isoprotenerol, with beating rates reaching up to 560 bpm. iSAB/Iso: n = 9; iSAB/Carb: n = 5. The data represent seven independent experiments and are presented as means ± SD. See also Figures S1 and S2 .
    Figure Legend Snippet: Electrophysiological Parameters of Pacemaker-like Cardiomyocytes Obtained from iSABs (A) Distribution of pacemaker cells as shown by single-cell patch-clamp and funny-channel measurements: > 81% of pacemaker-like cells were achieved; 19% of these represented immature pacemaker cells and the rest were mature pacemaker cells (n = 65). The data represent seven independent experiments and are presented as means ± SD. See also Tables S1–S3 . (B) Patch-clamp protocol. Voltage was applied to elicit the hyperpolarization-activated current for I f recordings from isolated iSAB cells. (C) Example of I f current, recorded from an isolated iSAB-derived cell. (D and E) Current density (D) and time constant of activation (E) at –130 mV, demonstrating a robust I f expression with slow activation kinetics typical of the HCN4 channel subtype and mature SA nodal I f (n = 17). The data represent three independent experiments and are presented as means ± SD. (F) Reaction to β-adrenergic (isoprotenerol) and muscarinic (carbachol) stimulation leads to typical accelerated versus decelerated AP rates. iSAB-derived cells show a pronounced response to isoprotenerol, with beating rates reaching up to 560 bpm. iSAB/Iso: n = 9; iSAB/Carb: n = 5. The data represent seven independent experiments and are presented as means ± SD. See also Figures S1 and S2 .

    Techniques Used: Patch Clamp, Isolation, Derivative Assay, Activation Assay, Expressing

    5) Product Images from "The sinus venosus myocardium contributes to the atrioventricular canal: potential role during atrioventricular node development?"

    Article Title: The sinus venosus myocardium contributes to the atrioventricular canal: potential role during atrioventricular node development?

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.12525

    Characterization of the myocardial continuity between the sinus venosus and AV canal. (A) Whole mount ISL1 staining, posterior view of the heart. Arrows indicate ISL1+ cells in continuity sinus venosus and AV canal. Rectangle 1 shows region from which SV-AVC continuity cells were collected. Rectangle 2 shows origin of right ventricular cells for patch clamp. (B–E) mRNA expression after laser capture microscopy (LCM) of TNNT2, NKX2-5, HCN4 and ISL1 . (F–M) Results of single cell patch-clamp experiments. (F and J) Representative tracing of SV-AVC continuity cells (F, pacemaker-like tracing) and RV cells (J, ventricular-like phenotype). (G–I, K–M) Graphs showing the difference in maximum upstroke velocity (G), frequency (H), amplitude (I), APD50 (K), APD90 (L) and maximum diastolic potential (M). * Indicates P
    Figure Legend Snippet: Characterization of the myocardial continuity between the sinus venosus and AV canal. (A) Whole mount ISL1 staining, posterior view of the heart. Arrows indicate ISL1+ cells in continuity sinus venosus and AV canal. Rectangle 1 shows region from which SV-AVC continuity cells were collected. Rectangle 2 shows origin of right ventricular cells for patch clamp. (B–E) mRNA expression after laser capture microscopy (LCM) of TNNT2, NKX2-5, HCN4 and ISL1 . (F–M) Results of single cell patch-clamp experiments. (F and J) Representative tracing of SV-AVC continuity cells (F, pacemaker-like tracing) and RV cells (J, ventricular-like phenotype). (G–I, K–M) Graphs showing the difference in maximum upstroke velocity (G), frequency (H), amplitude (I), APD50 (K), APD90 (L) and maximum diastolic potential (M). * Indicates P

    Techniques Used: Staining, Patch Clamp, Expressing, Microscopy, Laser Capture Microdissection

    HCN4 expression in myocardial continuity between sinus venosus and posterior AV canal. (A) TNNI2 stain of HH19 chick embryo. Arrow indicates continuity between sinus venosus myocardium and posterior AV canal. (B) ISH shows HCN4 mRNA expression in continuity (arrow). (C) Merge of ISL1, TNNI2, HCN4 and DAPI staining of E11.5 mouse embryo, showing the myocardial continuity between the sinus venosus and posterior AV canal. The ISL1+/HCN4+/TNNI2+ SAN (arrowhead) and RVV (arrow) are shown. Note the thick endocardial cushions (asterisks) in the AVC. (D–F) Higher magnifications of boxed area in C and D. ISL1+/TNNI2+/HCN4+ cells are seen in the continuity between sinus venosus myocardium and posterior AV canal myocardium. (E and F) Grey values of ISL1 (E) and HCN4 (F). AVC: atrioventricular canal; LA: left atrium; LB: long bud; LCV: left cardinal vein; LV: left ventricle; OFT: outflow tract; RA: right atrium; RCV: right cardinal vein; SV: sinus venosus; V: ventricle. (C–F) White: TNNI2, green: ISL1, red: HCN4, blue: DAPI; scale bars: 50 μm.
    Figure Legend Snippet: HCN4 expression in myocardial continuity between sinus venosus and posterior AV canal. (A) TNNI2 stain of HH19 chick embryo. Arrow indicates continuity between sinus venosus myocardium and posterior AV canal. (B) ISH shows HCN4 mRNA expression in continuity (arrow). (C) Merge of ISL1, TNNI2, HCN4 and DAPI staining of E11.5 mouse embryo, showing the myocardial continuity between the sinus venosus and posterior AV canal. The ISL1+/HCN4+/TNNI2+ SAN (arrowhead) and RVV (arrow) are shown. Note the thick endocardial cushions (asterisks) in the AVC. (D–F) Higher magnifications of boxed area in C and D. ISL1+/TNNI2+/HCN4+ cells are seen in the continuity between sinus venosus myocardium and posterior AV canal myocardium. (E and F) Grey values of ISL1 (E) and HCN4 (F). AVC: atrioventricular canal; LA: left atrium; LB: long bud; LCV: left cardinal vein; LV: left ventricle; OFT: outflow tract; RA: right atrium; RCV: right cardinal vein; SV: sinus venosus; V: ventricle. (C–F) White: TNNI2, green: ISL1, red: HCN4, blue: DAPI; scale bars: 50 μm.

    Techniques Used: Expressing, Staining, In Situ Hybridization

    Related Articles

    Incubation:

    Article Title: Cohesin-protein Shugoshin-1 controls cardiac automaticity via HCN4 pacemaker channel
    Article Snippet: .. They were then incubated with rabbit polyclonal anti-HCN4 antibody (1:200; APC-052, Alomone Labs) mixed with mouse monoclonal anti-SGO1 (1:200; ab58023, Abcam) in 2% normal goat serum PBS at 4 °C overnight. ..

    Immunolabeling:

    Article Title: Presynaptic HCN channels constrain GABAergic synaptic transmission in pyramidal cells of the medial prefrontal cortex
    Article Snippet: .. Sequential primary immunolabeling for HCN1, HCN2 or HCN4 was performed using anti-HCN1, HCN2 or HCN4 rabbit antibodies (1:40; Alomone Laboratories, Israel,. ..

    other:

    Article Title: Structural Identification of the Pacemaker Cells and Expression of Hyperpolarization-Activated Cyclic Nucleotide-Gated (HCN) Channels in the Heart of the Wild Atlantic Cod, Gadus morhua (Linnaeus, 1758)
    Article Snippet: HCN4 antibodies as reliable markers of the pacemaker cells as well as the controls of HCN4 and Islet-1 antibodies in the heart of zebrafish were reported by [ , ].

    Expressing:

    Article Title: The HCN Channel Blocker ZD7288 Induces Emesis in the Least Shrew (Cryptotis parva)
    Article Snippet: .. Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels ExpressionTo analyze the expression and distribution of HCN channel subtypes 1–4 in the least shrew brainstem dorsal vagal complex area, the following primary and secondary antibodies were used: rabbit anti-HCN1 antibody (1:300; ab229340, Abcam), mouse anti-HCN2 antibody (1:300; ab84817, Abcam), mouse anti-HCN3 antibody (1:300; ab84818, Abcam), rabbit anti-HCN4 antibody (1:300; APC-052, Alomone Labs), Alexa Fluor 594 donkey anti-rabbit (1:500, Abcam) and 488 conjugated donkey anti-mouse secondary antibodies (1:500, Invitrogen). ..

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    Alomone Labs hcn4
    Transcriptional profiling of Cx30.2-lacZ + cells identifies MyoR as an AVC-enriched transcript. (A) Strategy used for purifying E12.5 Cx30.2-lacZ + AVC cells by flow cytometry. (B) Partial list of genes found to be enriched in microarray analysis. Fold enrichment refers to transcript levels in Cx30.2-lacZ + cells relative to lacZ -null cells. Note that MyoR transcripts were 7-fold more abundant in Cx30.2-lacZ + cells within the E12.5 heart. (C) qRT-PCR analysis of microdissected E10.5 AVC tissue demonstrating enrichment of MyoR. Fold enrichment refers to transcript levels in dissected AVC tissue versus whole heart. (D) Developmental analysis of MyoR expression by qRT-PCR analysis. MyoR expression in the whole heart at a particular time point relative to E10.5 is shown. MyoR transcripts were found to peak during late gestation (E16.5). (E) qRT-PCR analysis of MyoR expression level in Cx30.2-EGFP + AVC cells isolated at E16.5. MyoR expression is shown relative to transcript levels in EGFP − AVC cells. (F) Western blot analysis of AVC and ventricular nuclear extracts dissected from E16.5 embryos (top) with the indicated antibodies. Band quantitation and normalization demonstrated ∼14-fold enrichment of MyoR in the E16.5 AVC relative to ventricular tissue. (G) Immunostaining of consecutive sections through an E16.5 Cx30.2-EGFP heart for GFP, Gata4, <t>Hcn4,</t> and MyoR, demonstrating localization of Cx30.2, Gata4, and MyoR to the Hcn4 + AVN (dashed outline). All sections were counterstained with DAPI to stain cell nuclei. Scale bar, 20 μm. LV, left ventricle; RV, right ventricle; FDG, fluorescein digalactopyranoside; AVC, atrioventricular canal; AVN, atrioventricular node.
    Hcn4, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hcn4/product/Alomone Labs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hcn4 - by Bioz Stars, 2021-09
    95/100 stars
      Buy from Supplier

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    Transcriptional profiling of Cx30.2-lacZ + cells identifies MyoR as an AVC-enriched transcript. (A) Strategy used for purifying E12.5 Cx30.2-lacZ + AVC cells by flow cytometry. (B) Partial list of genes found to be enriched in microarray analysis. Fold enrichment refers to transcript levels in Cx30.2-lacZ + cells relative to lacZ -null cells. Note that MyoR transcripts were 7-fold more abundant in Cx30.2-lacZ + cells within the E12.5 heart. (C) qRT-PCR analysis of microdissected E10.5 AVC tissue demonstrating enrichment of MyoR. Fold enrichment refers to transcript levels in dissected AVC tissue versus whole heart. (D) Developmental analysis of MyoR expression by qRT-PCR analysis. MyoR expression in the whole heart at a particular time point relative to E10.5 is shown. MyoR transcripts were found to peak during late gestation (E16.5). (E) qRT-PCR analysis of MyoR expression level in Cx30.2-EGFP + AVC cells isolated at E16.5. MyoR expression is shown relative to transcript levels in EGFP − AVC cells. (F) Western blot analysis of AVC and ventricular nuclear extracts dissected from E16.5 embryos (top) with the indicated antibodies. Band quantitation and normalization demonstrated ∼14-fold enrichment of MyoR in the E16.5 AVC relative to ventricular tissue. (G) Immunostaining of consecutive sections through an E16.5 Cx30.2-EGFP heart for GFP, Gata4, Hcn4, and MyoR, demonstrating localization of Cx30.2, Gata4, and MyoR to the Hcn4 + AVN (dashed outline). All sections were counterstained with DAPI to stain cell nuclei. Scale bar, 20 μm. LV, left ventricle; RV, right ventricle; FDG, fluorescein digalactopyranoside; AVC, atrioventricular canal; AVN, atrioventricular node.

    Journal: Molecular and Cellular Biology

    Article Title: MyoR Modulates Cardiac Conduction by Repressing Gata4

    doi: 10.1128/MCB.00860-14

    Figure Lengend Snippet: Transcriptional profiling of Cx30.2-lacZ + cells identifies MyoR as an AVC-enriched transcript. (A) Strategy used for purifying E12.5 Cx30.2-lacZ + AVC cells by flow cytometry. (B) Partial list of genes found to be enriched in microarray analysis. Fold enrichment refers to transcript levels in Cx30.2-lacZ + cells relative to lacZ -null cells. Note that MyoR transcripts were 7-fold more abundant in Cx30.2-lacZ + cells within the E12.5 heart. (C) qRT-PCR analysis of microdissected E10.5 AVC tissue demonstrating enrichment of MyoR. Fold enrichment refers to transcript levels in dissected AVC tissue versus whole heart. (D) Developmental analysis of MyoR expression by qRT-PCR analysis. MyoR expression in the whole heart at a particular time point relative to E10.5 is shown. MyoR transcripts were found to peak during late gestation (E16.5). (E) qRT-PCR analysis of MyoR expression level in Cx30.2-EGFP + AVC cells isolated at E16.5. MyoR expression is shown relative to transcript levels in EGFP − AVC cells. (F) Western blot analysis of AVC and ventricular nuclear extracts dissected from E16.5 embryos (top) with the indicated antibodies. Band quantitation and normalization demonstrated ∼14-fold enrichment of MyoR in the E16.5 AVC relative to ventricular tissue. (G) Immunostaining of consecutive sections through an E16.5 Cx30.2-EGFP heart for GFP, Gata4, Hcn4, and MyoR, demonstrating localization of Cx30.2, Gata4, and MyoR to the Hcn4 + AVN (dashed outline). All sections were counterstained with DAPI to stain cell nuclei. Scale bar, 20 μm. LV, left ventricle; RV, right ventricle; FDG, fluorescein digalactopyranoside; AVC, atrioventricular canal; AVN, atrioventricular node.

    Article Snippet: Consecutive sections through the AVN were stained with GFP (Life Technologies, Grand Island, NY), Gata4 (Santa Cruz Biotechnology, Dallas, TX), MyoR (Santa Cruz Biotechnology), and Hcn4 (Alomone Labs, Jerusalem, Israel) primary antibodies, followed by detection with Alexa Fluor-labeled secondary antibodies (Life Technologies) according to the manufacturer's suggested protocol.

    Techniques: Flow Cytometry, Cytometry, Microarray, Quantitative RT-PCR, Expressing, Isolation, Western Blot, Quantitation Assay, Immunostaining, Staining