rat circrna array  (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: To investigate the circRNA expression profile of DCM, we have applied the Arraystar Rat circRNA Array analysis of specimen from three DCM samples and three normal controls.

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: To investigate the circRNA expression profile of DCM, we have applied the Arraystar Rat circRNA Array analysis of specimen from three DCM samples and three normal controls.

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: To investigate the circRNA expression profile of DCM, we have applied the Arraystar Rat circRNA Array analysis of specimen from three DCM samples and three normal controls.

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: To investigate the circRNA expression profile of DCM, we have applied the Arraystar Rat circRNA Array analysis of specimen from three DCM samples and three normal controls.

Techniques: Western Blot, Expressing, Transfection

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

Journal: Frontiers in Neuroscience

Article Title: Identification and characterization of N6-methyladenosine circular RNAs in the spinal cord of morphine-tolerant rats

doi: 10.3389/fnins.2022.967768

Figure Lengend Snippet: The animal model and the overall features of m 6 A-circRNA in morphine-tolerant rats. (A) Morphine-induced antinociception was assessed through the tail-flick test, and tail-flick latency was converted to %MPE ( n = 4), *** P < 0.001 compared with the NS group using Two-way repeated measures of ANOVA followed by Bonferroni’s post-hoc test. (B) Expression of RNA demethylase and methyltransferase determined by qRT-PCR. ( n = 4) ** P < 0.01, *** P < 0.001 compared with the NS group using a two-tailed unpaired Student’s t -test. (C) Expression of RNA methylase determined by qRT-PCR ( n = 4). * P < 0.05 compared with the NS group using a two-tailed unpaired Student’s t -test. (D) Hierarchical clustering shows global m 6 A-methylated circRNAs between the MT group and NS group ( n = 4).

Article Snippet: The labeled cRNAs were combined and hybridized to Arraystar Rat CircRNA Epitranscriptomic Arrays (8x15K, Arraystar) at 65°C for 17 h in an Agilent Hybridization Oven.

Techniques: Animal Model, Tail Flick Test, Expressing, Quantitative RT-PCR, Two Tailed Test, Methylation

Journal: Frontiers in Neuroscience

Article Title: Identification and characterization of N6-methyladenosine circular RNAs in the spinal cord of morphine-tolerant rats

doi: 10.3389/fnins.2022.967768

Figure Lengend Snippet: The detailed information of significantly top 10 hypermethylated m 6 A-circRNA and top 10 hypomethylated m 6 A-circRNAs.

Article Snippet: The labeled cRNAs were combined and hybridized to Arraystar Rat CircRNA Epitranscriptomic Arrays (8x15K, Arraystar) at 65°C for 17 h in an Agilent Hybridization Oven.

Techniques:

Journal: Frontiers in Neuroscience

Article Title: Identification and characterization of N6-methyladenosine circular RNAs in the spinal cord of morphine-tolerant rats

doi: 10.3389/fnins.2022.967768

Figure Lengend Snippet: Joint analysis of m 6 A methylation and expression patterns of circRNAs in morphine tolerance. (A) The nine-quadrant graph shows the relationship between m 6 A quantity and the expression of circRNAs. (B) Cumulative distribution of circRNAs expression between MT group and NS group for m 6 A-circRNAs (blue) and non-m 6 A- circRNAs (red). (C) The upset diagram shows the m 6 A quantity, expression, and m 6 A level of circRNAs and presents 14 different methylation and expression patterns of circRNAs. (D) The dot plot shows the distribution of 96 circRNA with both dysregulated m 6 A quantity and m 6 A level. The orange dots (hyper-up) show 27 circRNAs were hypermethylated and up-expressed, 2 navy blue dots (hypo-down) represent circRNAs were hypomethylated and down-expressed; 1 black dot (hypo-up) represent circRNA were hypomethylated and up-expressed; the light blue dots (hypo-no) represent 66 circRNAs with decreased m6A modification quantity but unchanged expression levels.

Article Snippet: The labeled cRNAs were combined and hybridized to Arraystar Rat CircRNA Epitranscriptomic Arrays (8x15K, Arraystar) at 65°C for 17 h in an Agilent Hybridization Oven.

Techniques: Methylation, Expressing, Modification

Journal: Frontiers in Neuroscience

Article Title: Identification and characterization of N6-methyladenosine circular RNAs in the spinal cord of morphine-tolerant rats

doi: 10.3389/fnins.2022.967768

Figure Lengend Snippet: Construction of circRNA–miRNA networks in morphine tolerance using circRNAs with differences in both m 6 A quantity and m 6 A modification level.

Article Snippet: The labeled cRNAs were combined and hybridized to Arraystar Rat CircRNA Epitranscriptomic Arrays (8x15K, Arraystar) at 65°C for 17 h in an Agilent Hybridization Oven.

Techniques: Modification

Journal: Frontiers in Neuroscience

Article Title: Identification and characterization of N6-methyladenosine circular RNAs in the spinal cord of morphine-tolerant rats

doi: 10.3389/fnins.2022.967768

Figure Lengend Snippet: M 6 A levels of the four circRNAs involved in the circRNA–miRNA networks were verified by MeRIP-qPCR. Relative m 6 A methylation level was calculated as the percentage of modified transcripts in all transcripts. P -values were calculated using Student’s t -test ( n = 4). ** P < 0.01, *** P < 0.001 compared with the NS group using a two-tailed unpaired Student’s t -test.

Article Snippet: The labeled cRNAs were combined and hybridized to Arraystar Rat CircRNA Epitranscriptomic Arrays (8x15K, Arraystar) at 65°C for 17 h in an Agilent Hybridization Oven.

Techniques: Methylation, Modification, Two Tailed Test