mouse cardiomyocytes  (Servicebio Inc)

Journal: Journal of Inflammation Research

Article Title: Protective Effects of CR6-Interacting Factor 1 Against Angiotensin II-Induced Atrial Fibrillation by Regulating the SIRT1/eNOS Signaling Pathway and Cardiomyocyte Remodeling

doi: 10.2147/JIR.S596132

Figure Lengend Snippet: CRIF1 was downregulated in the AF mice. ( A ) Atrial tissue of mice was stained with hematoxylin and eosin (H&E) to display the morphological structure. CRIF1 expression in the atrial tissue of mice was detected using immunohistochemistry (200×). ( B ) The CRIF1 mRNA levels in the atrial tissue were detected by RT-qPCR. The mice cardiac muscle cell line HL-1 was treated with Ang II (0, 0.1, 0.2, 0.5, 1 and 2 μM) for 24 h. ( C ) The CRIF1 mRNA expression was determined by RT-qPCR. Data are presented as mean ± SD (n=8 mice in each group). t -test was applied. ***P<0.001 vs control group.

Article Snippet: The mouse cardiac muscle cell line HL-1 was purchased from Procell (CL-0605, China).

Techniques: Staining, Expressing, Immunohistochemistry, Quantitative RT-PCR, Control

Journal: Journal of Inflammation Research

Article Title: Protective Effects of CR6-Interacting Factor 1 Against Angiotensin II-Induced Atrial Fibrillation by Regulating the SIRT1/eNOS Signaling Pathway and Cardiomyocyte Remodeling

doi: 10.2147/JIR.S596132

Figure Lengend Snippet: Overexpression of CRIF1 activates the SIRT1/eNOS pathway in cardiomyocytes treated with Ang II. HL-1 cells were transfected with pcDNA3.1-CRIF1 (OE-CRIF1) or vector for 24 h, and then treated with Ang II (1 μM) for an additional 24 h. The mRNA expression levels of ( A ) CRIF1, ( B ) SIRT1 and ( C ) eNOS were evaluated using RT-qPCR. HL-1 cells were transfected with pcDNA3.1-CRIF1 (OE-CRIF1) or vector for 24 h, and were then cotreated with a SIRT1 inhibitor EX527 (10 μM) and Ang II (1 μM) for further 24 h. ( D ) The cell apoptosis was assessed by staining with TUNEL and DAPI, and observed under a fluorescence microscope (200 X). ( E ) Apoptosis was quantified by calculating TUNEL positive cells (normalized to DAPI-stained cells). Data are presented as mean ± SD in triplicates. ***P<0.001 vs control group; ### P<0.001 vs Ang II+Vector group; $$$ P<0.001 vs OE-CRIF1 group.

Article Snippet: The mouse cardiac muscle cell line HL-1 was purchased from Procell (CL-0605, China).

Techniques: Over Expression, Transfection, Plasmid Preparation, Expressing, Quantitative RT-PCR, Staining, TUNEL Assay, Fluorescence, Microscopy, Control

Journal: Journal of Inflammation Research

Article Title: Protective Effects of CR6-Interacting Factor 1 Against Angiotensin II-Induced Atrial Fibrillation by Regulating the SIRT1/eNOS Signaling Pathway and Cardiomyocyte Remodeling

doi: 10.2147/JIR.S596132

Figure Lengend Snippet: CRIF1 overexpression inhibits Ang II–induced hypertrophy of HL-1 cells. ( A ) Cells were stained with α-actinin antibody and observed under a fluorescence microscope. Representative images are shown (200×). RT-qPCR was used to assess the mRNA expression levels of hypertrophy-related genes, including ( B ) ANP, ( C ) BNP, and ( D ) β-MHC. Data are presented as mean ± SD in triplicate. ***P<0.001 vs control group; ### P<0.001 vs Ang II+Vector group; $$$ P<0.001 vs OE-CRIF1 group.

Article Snippet: The mouse cardiac muscle cell line HL-1 was purchased from Procell (CL-0605, China).

Techniques: Over Expression, Staining, Fluorescence, Microscopy, Quantitative RT-PCR, Expressing, Control, Plasmid Preparation

Journal: Journal of Inflammation Research

Article Title: Protective Effects of CR6-Interacting Factor 1 Against Angiotensin II-Induced Atrial Fibrillation by Regulating the SIRT1/eNOS Signaling Pathway and Cardiomyocyte Remodeling

doi: 10.2147/JIR.S596132

Figure Lengend Snippet: CRIF1 overexpression suppresses Ang II–induced inflammation in cardiomyocytes. ( A – C ) RT-qPCR was used to determine the mRNA levels of the pro-inflammatory cytokines TNF-α, IL-1β, and IL-6. ( D – F ) ELISA was used to measure the levels of TNF-α, IL-1β, and IL-6 in the culture medium of HL-1 cells. Data are presented as mean ± SD in triplicate. ***P<0.001 vs control group; ### P<0.001 vs Ang II+Vector group; $$$ P<0.001 vs OE-CRIF1 group.

Article Snippet: The mouse cardiac muscle cell line HL-1 was purchased from Procell (CL-0605, China).

Techniques: Over Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control, Plasmid Preparation

Journal: Journal of Inflammation Research

Article Title: Protective Effects of CR6-Interacting Factor 1 Against Angiotensin II-Induced Atrial Fibrillation by Regulating the SIRT1/eNOS Signaling Pathway and Cardiomyocyte Remodeling

doi: 10.2147/JIR.S596132

Figure Lengend Snippet: Overexpression of CRIF1 inhibits intracellular ROS generation and oxidative stress in cardiomyocytes treated with Ang II. ( A ) Cells were stained with DHE, and representative images of intracellular ROS are shown (200×). ( B ) The extent of intracellular ROS was quantified by counting DHE-positive cells (normalized to DAPI-stained cells). Cell lysates of HL-1 cells were used to detect the oxidative stress markers: ( C ) MDA, ( D ) SOD, ( E ) CAT, and ( F ) NO. Data are presented as mean ± SD in triplicate. ***P<0.001 vs control group; ### P<0.001 vs Ang II+Vector group; $$$ P<0.001 vs OE-CRIF1 group.

Article Snippet: The mouse cardiac muscle cell line HL-1 was purchased from Procell (CL-0605, China).

Techniques: Over Expression, Staining, Control, Plasmid Preparation

Journal: Europace

Article Title: Fto-mediated m 6 A demethylation of Lox drives atrial fibrosis and promotes atrial fibrillation in a murine model of hyperthyroidism

doi: 10.1093/europace/euag047

Figure Lengend Snippet: Cardiomyocyte-specific deletion of Fto alleviated T4-induced AF. ( A ) Representative traces of AF induced by electrical stimulation in Myh6-cre − ; Fto fl/fl mice and Myh6-cre + ; Fto fl/fl mice in saline and T4 groups. ( B ) Quantification of AF inducibility ( n = 10). ( C ) AF duration was quantified only in animals with inducible episodes. The sample size for each group is n = 10, of which 2 (Myh6-cre − ; Fto fl/fl ) and 7 (Myh6-cre + ; Fto fl/fl ) animals did not have inducible AF and are therefore not included in (C) . ( D ) Quantification of HW/TL ( n = 10). ( E ) Representative maps of atrial activation time and dispersion of conduction in Myh6-cre − ; Fto fl/fl mice and Myh6-cre + ; Fto fl/fl mice in saline and T4 groups ( n = 6). ( F ) Quantification of active time ( n = 6). ( G ) Quantification of dispersion of conduction ( n = 6). ( H ) Quantification of LACV ( n = 6). ( I ) Quantification of ERP ( n = 6). ( J ) Representative images of LAD and Masson staining in atria. ( K ) Quantification of fibrosis area ( n = 6). ( L ) Quantification of LAD ( n = 6). AF, atrial fibrillation; β-Actin, beta-actin; ERP, effective refractory period; Fto, fat mass and obesity-associated protein; HR, heart rate; HW/TL, heart weight-to-tibial length ratio; LACV, left atrial conduction velocity; LAD, left atrial diameter. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant. Statistical analysis was performed with ordinary one-way ANOVA ( D , F , G , H , I , K , and L ), Fisher's exact test ( B ) and unpaired Student's t -test ( C ).

Article Snippet: Cardiomyocyte-specific Fto knockout mice ( Myh6-Cre + ; Fto fl/fl ) were established on a C57BL/6J background (Shanghai Model Organisms, NM-CKO-190005) and analysed at 2 months of age.

Techniques: Saline, Activation Assay, Dispersion, Staining

Journal: Europace

Article Title: Fto-mediated m 6 A demethylation of Lox drives atrial fibrosis and promotes atrial fibrillation in a murine model of hyperthyroidism

doi: 10.1093/europace/euag047

Figure Lengend Snippet: Cardiomyocyte-specific Fto knockout attenuated Lox expression in T4-treated mice. ( A and B ) GO functional enrichment analysis of differentially expressed genes. ( C ) Venn diagram showing overlap of differentially expressed genes between two groups. ( D ) Representative immunoblots of Fto and Lox protein in Myh6-cre − ; Fto fl/fl mice and Myh6-cre + ; Fto fl/fl mice in saline and T4 groups ( n = 6). ( E ) Quantification of Lox mRNA levels ( n = 6). ( F ) Quantification of Fto protein levels ( n = 6). ( G ) Quantification of Lox protein levels ( n = 6). β-Actin, beta-actin; Fto, fat mass and obesity-associated protein; GO, gene ontology; Lox, lysyl oxidase. **** P < 0.0001, comparison made using ordinary one-way ANOVA.

Article Snippet: Cardiomyocyte-specific Fto knockout mice ( Myh6-Cre + ; Fto fl/fl ) were established on a C57BL/6J background (Shanghai Model Organisms, NM-CKO-190005) and analysed at 2 months of age.

Techniques: Knock-Out, Expressing, Functional Assay, Western Blot, Saline, Comparison

Journal: Europace

Article Title: Fto-mediated m 6 A demethylation of Lox drives atrial fibrosis and promotes atrial fibrillation in a murine model of hyperthyroidism

doi: 10.1093/europace/euag047

Figure Lengend Snippet: Cardiomyocyte-specific Fto overexpression promoted AF susceptibility via m 6 A-dependent mechanisms. ( A ) Representative immunoblots of Fto and Lox proteins in mice injected with AAV9-cTnT- Fto wt -oe, AAV9-cTnT- Fto mut -oe, and AAV9-cTnT-nc ( n = 6). ( B and C ) Quantification of Fto and Lox protein levels ( n = 6). ( D ) Representative intracardiac bipolar electrograms showing induction of AF from mice injected with AAV9-cTnT- Fto wt -oe, AAV9-cTnT- Fto mut -oe, and AAV9-cTnT-nc. ( E ) Quantification of AF inducibility ( n = 10). ( F ) AF was inducible only in the AAV9-cTnT- Fto wt -oe group (7 out of 10 animals). Therefore, data are presented only for this group. AF was not inducible in any animal from the AAV9-cTnT-nc ( n = 10) or AAV9-cTnT- Fto mut -oe ( n = 10) groups; hence, no AF duration data are available for these groups, and they are not represented in this graph. ( G ) Representative maps of atrial activation time and dispersion of conduction. ( H ) Quantification of active time ( n = 6). ( I ) Quantification of dispersion of conduction ( n = 6). ( J ) Quantification of LACV ( n = 6). ( K ) Quantification of ERP ( n = 6). ( L ) Representative images of LAD and Masson staining. ( M ) Quantification of LAD ( n = 6). ( N ) Quantification of percentage of fibrotic area ( n = 6). AF, atrial fibrillation; β-Actin, beta-actin; ERP, effective refractory period; Fto, fat mass and obesity-associated protein; ; LACV, left atrial conduction velocity; LAD, left atrial diameter; Lox, lysyl oxidase. ** P < 0.01; *** P < 0.001; **** P < 0.0001. Statistical analysis was performed with ordinary one-way ANOVA ( B , C , H , I , J , K , M , and N ), Fisher's exact test ( E ).

Article Snippet: Cardiomyocyte-specific Fto knockout mice ( Myh6-Cre + ; Fto fl/fl ) were established on a C57BL/6J background (Shanghai Model Organisms, NM-CKO-190005) and analysed at 2 months of age.

Techniques: Over Expression, Western Blot, Injection, Activation Assay, Dispersion, Staining