rat circrna array v2.0 (8 × 15 k  (Arraystar inc)

Journal: Journal of Diabetes

Article Title: CircMAP3K5 promotes cardiomyocyte apoptosis in diabetic cardiomyopathy by regulating miR ‐22‐3p/ DAPK2 Axis

doi: 10.1111/1753-0407.13471

Figure Lengend Snippet: Differential expression of circRNA by A, scatter plot, B, volcanic map, and C, cluster analysis in normal group and diabetic cardiomyopathy (DCM) group. circRNA, circular RNA.

Article Snippet: These circRNAs were amplified and transcribed into fluorescent cRNA using a random primer method (Arraystar Super RNA Labeling Kit; Arraystar). labeled cRNAs that were hybridized to Arraystar Rat circRNA Array V2.0 (8 × 15 K) with a total of 14 145 circRNA probes on the chip.

Techniques: Expressing

Journal: Journal of Diabetes

Article Title: CircMAP3K5 promotes cardiomyocyte apoptosis in diabetic cardiomyopathy by regulating miR ‐22‐3p/ DAPK2 Axis

doi: 10.1111/1753-0407.13471

Figure Lengend Snippet: Depletion of circRNA Mitogen‐activated protein kinase kinase kinase 5 (circMAP3K5) attenuates cardiomyocyte apoptosis in a diabetic cardiomyopathy (DCM) cell model. A, quantitative real‐time polymerase chain reaction results show expression levels of circMAP3K5 in samples of DCM ( n = 3) and control ( n = 3) cells. B, Western blots showing expression levels of Bcl‐2, Bax, and cleaved caspase‐3 proteins in the samples. C, Flow cytometry results depict the rate of apoptosis. Statistically significant difference: * p < .05 ( n = 3). circRNA, circular RNA; HG, high glucose.

Article Snippet: These circRNAs were amplified and transcribed into fluorescent cRNA using a random primer method (Arraystar Super RNA Labeling Kit; Arraystar). labeled cRNAs that were hybridized to Arraystar Rat circRNA Array V2.0 (8 × 15 K) with a total of 14 145 circRNA probes on the chip.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Flow Cytometry

Journal: Journal of Diabetes

Article Title: CircMAP3K5 promotes cardiomyocyte apoptosis in diabetic cardiomyopathy by regulating miR ‐22‐3p/ DAPK2 Axis

doi: 10.1111/1753-0407.13471

Figure Lengend Snippet: CircRNA mitogen‐activated protein kinase kinase kinase 5 (circMAP3K5) acts as a sponge for miR‐22‐3p in cardiomyocytes. A, Predicted potential interactions between circMAP3K5 and miR‐22‐3p according to Miranda software. B, quantitative real‐time polymerase chain reaction (qRT‐PCR) results showing expression level of miR‐22‐3p in H9c2 cells. C, Luciferase activity in wild‐type circMAP3K5 (circMAP3K5 WT) and a mutant with a miR‐22‐3p binding site (circMAP3K5 MUT) in 293 T cells transfected with a control mimic or a miR‐22‐3p mimic. D, qRT‐PCR results depicting levels of miR‐22‐3p expression in H9c2 cells transfected with either si‐MAP3K5 NC or si‐MAP3K5. E, qRT‐PCR results showing levels of miR‐22‐3p expression in H9c2 cells transfected with either pLc‐MAP3K5 vector or pLc‐MAP3K5. F, Western blots showing expression levels of the apoptosis regulatory protein. Statistically significant difference: * p < .05 ( n = 3). HG, high glucose; miRNA, microRNA.

Article Snippet: These circRNAs were amplified and transcribed into fluorescent cRNA using a random primer method (Arraystar Super RNA Labeling Kit; Arraystar). labeled cRNAs that were hybridized to Arraystar Rat circRNA Array V2.0 (8 × 15 K) with a total of 14 145 circRNA probes on the chip.

Techniques: Software, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Luciferase, Activity Assay, Mutagenesis, Binding Assay, Transfection, Plasmid Preparation, Western Blot

Journal: Journal of Diabetes

Article Title: CircMAP3K5 promotes cardiomyocyte apoptosis in diabetic cardiomyopathy by regulating miR ‐22‐3p/ DAPK2 Axis

doi: 10.1111/1753-0407.13471

Figure Lengend Snippet: CircRNA mitogen‐activated protein kinase kinase kinase 5 (circMAP3K5) promotes HG‐induced cardiomyocyte apoptosis by regulating the miR‐22‐3p/DAPK2 axis. A, Western blots showing expression levels of DAPK2 protein in cardiac myocytes transfected with either si‐circMAP3K5 or pLcMAP3K5, and exposed to high glucose concentrations (25 mmol/L) for 48 h. B, Western blots showing expression levels of apoptosis‐related proteins in cardiomyocytes transfected with si‐circMAP3K5, miR‐22‐3p inhibitor, and/or pcDNA‐DAPK2, under high glucose concentrations (25 mmol/L). C, Proposed mechanism of circMAP3K5 mediated DCM pathological process: Increases circMAP3K5 levels in the diabetic cardiomyopathy (DCM) heart leading to its interaction with miR‐22, which affects cardiomyocytes apoptosis through upregulation of DAPK2. DAPK2, death‐associated protein kinase 2; HG, high glucose.

Article Snippet: These circRNAs were amplified and transcribed into fluorescent cRNA using a random primer method (Arraystar Super RNA Labeling Kit; Arraystar). labeled cRNAs that were hybridized to Arraystar Rat circRNA Array V2.0 (8 × 15 K) with a total of 14 145 circRNA probes on the chip.

Techniques: Western Blot, Expressing, Transfection