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CircCacna1c competitively binds to miR-29b-2-5p. ( a ) Target miRNAs of circCacna1c predicated by Arraystar’s homemade miRNA target prediction software based on <t>TargetScan</t> and miRanda. ( b ) The expression of miR-29b-2-5p and miR-669a-5p in ISO-induced heart tissue. T -test was used for statistical analysis, n = 3–5. ( c ) Potential binding sites of miR-29b-2-5p on the circCacna1c sequence. ( d ) The expression of miR-29b-2-5p in H9c2 cells after knockdown of circCacna1c with/without ISO treatment. One-way ANOVA was used for statistical analysis, n = 3. ( e ) Verification of miR-29b-2-5p as sponged targets of circCacna1c by dual-luciferase reporter assay. Left: mutated nucleotides on the binding sites of circCacna1c with miR-29b-2-5p (shown as red letters); Right: the statistical chart of relative luciferase activity. T -test was used for statistical analysis, n = 3. Data are shown as mean ± SEM. * p < 0.05, ** p < 0.01. ISO: isoprenaline hydrochloride; WT: wild type; MUT: mutant.
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CircCacna1c competitively binds to miR-29b-2-5p. ( a ) Target miRNAs of circCacna1c predicated by Arraystar’s homemade miRNA target prediction software based on TargetScan and miRanda. ( b ) The expression of miR-29b-2-5p and miR-669a-5p in ISO-induced heart tissue. T -test was used for statistical analysis, n = 3–5. ( c ) Potential binding sites of miR-29b-2-5p on the circCacna1c sequence. ( d ) The expression of miR-29b-2-5p in H9c2 cells after knockdown of circCacna1c with/without ISO treatment. One-way ANOVA was used for statistical analysis, n = 3. ( e ) Verification of miR-29b-2-5p as sponged targets of circCacna1c by dual-luciferase reporter assay. Left: mutated nucleotides on the binding sites of circCacna1c with miR-29b-2-5p (shown as red letters); Right: the statistical chart of relative luciferase activity. T -test was used for statistical analysis, n = 3. Data are shown as mean ± SEM. * p < 0.05, ** p < 0.01. ISO: isoprenaline hydrochloride; WT: wild type; MUT: mutant.

Journal: Cells

Article Title: Silencing of circCacna1c Inhibits ISO-Induced Cardiac Hypertrophy through miR-29b-2-5p/NFATc1 Axis

doi: 10.3390/cells12121667

Figure Lengend Snippet: CircCacna1c competitively binds to miR-29b-2-5p. ( a ) Target miRNAs of circCacna1c predicated by Arraystar’s homemade miRNA target prediction software based on TargetScan and miRanda. ( b ) The expression of miR-29b-2-5p and miR-669a-5p in ISO-induced heart tissue. T -test was used for statistical analysis, n = 3–5. ( c ) Potential binding sites of miR-29b-2-5p on the circCacna1c sequence. ( d ) The expression of miR-29b-2-5p in H9c2 cells after knockdown of circCacna1c with/without ISO treatment. One-way ANOVA was used for statistical analysis, n = 3. ( e ) Verification of miR-29b-2-5p as sponged targets of circCacna1c by dual-luciferase reporter assay. Left: mutated nucleotides on the binding sites of circCacna1c with miR-29b-2-5p (shown as red letters); Right: the statistical chart of relative luciferase activity. T -test was used for statistical analysis, n = 3. Data are shown as mean ± SEM. * p < 0.05, ** p < 0.01. ISO: isoprenaline hydrochloride; WT: wild type; MUT: mutant.

Article Snippet: We then applied TargetScan and miRanda from Arraystar, which predicated five miRNAs that are targeted by circCacna1c, namely miR-135a-2-3p, miR-6914-5p, miR-5126, miR-29b-2-5p, and miR-669a-5p ( a and ).

Techniques: Software, Expressing, Binding Assay, Sequencing, Knockdown, Luciferase, Reporter Assay, Activity Assay, Mutagenesis

Silencing circCacna1c decreases hypertrophic gene expression via miR-29b-2-5p/NFATc1 axis. ( a ) Potential binding sites of miR-29b-2-5p on the NFATc1 sequence predicted by TargetScan. ( b ) Detection of the expression of NFATc1 mRNA in ISO-induced H9c2 cells by quantitative real-time polymerase chain reaction assay. ( c , d ) Detection of mRNA and protein expression of NFATc1 in H9c2 cells with overexpression of miR-29b-2-5p. T -test was used for statistical analysis. ( e , f ) Detection of the mRNA and protein expression of NFATc1 in ISO-induced H9c2 cells with overexpression of miR-29b-2-5p. ( g , h ) Detection of the mRNA and protein expression of NFATc1 in ISO-induced H9c2 cells with knockdown of circCacna1c. One-way ANOVA was used for statistical analysis. Data are shown as mean ± SEM. n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001. ISO: isoprenaline hydrochloride.

Journal: Cells

Article Title: Silencing of circCacna1c Inhibits ISO-Induced Cardiac Hypertrophy through miR-29b-2-5p/NFATc1 Axis

doi: 10.3390/cells12121667

Figure Lengend Snippet: Silencing circCacna1c decreases hypertrophic gene expression via miR-29b-2-5p/NFATc1 axis. ( a ) Potential binding sites of miR-29b-2-5p on the NFATc1 sequence predicted by TargetScan. ( b ) Detection of the expression of NFATc1 mRNA in ISO-induced H9c2 cells by quantitative real-time polymerase chain reaction assay. ( c , d ) Detection of mRNA and protein expression of NFATc1 in H9c2 cells with overexpression of miR-29b-2-5p. T -test was used for statistical analysis. ( e , f ) Detection of the mRNA and protein expression of NFATc1 in ISO-induced H9c2 cells with overexpression of miR-29b-2-5p. ( g , h ) Detection of the mRNA and protein expression of NFATc1 in ISO-induced H9c2 cells with knockdown of circCacna1c. One-way ANOVA was used for statistical analysis. Data are shown as mean ± SEM. n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001. ISO: isoprenaline hydrochloride.

Article Snippet: We then applied TargetScan and miRanda from Arraystar, which predicated five miRNAs that are targeted by circCacna1c, namely miR-135a-2-3p, miR-6914-5p, miR-5126, miR-29b-2-5p, and miR-669a-5p ( a and ).

Techniques: Gene Expression, Binding Assay, Sequencing, Expressing, Real-time Polymerase Chain Reaction, Over Expression, Knockdown