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Primers used in real-time <t> qPCR </t> analysis
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Primers used in real-time qPCR analysis

Journal: In Vitro Cellular & Developmental Biology. Animal

Article Title: Upregulation of miRNA-23a-3p rescues high glucose-induced cell apoptosis and proliferation inhibition in cardiomyocytes

doi: 10.1007/s11626-020-00518-6

Figure Lengend Snippet: Primers used in real-time qPCR analysis

Article Snippet: Real-time qPCR was performed with SuperReal PreMix Plus (FP 205, TianGen) and the miRcute Plus miRNA qPCR Detection Kit (FP 411, TianGen) on an Exicycler 96 PCR thermal block (Light Cycler 480, Roche, Basel, Switzerland).

Techniques: Sequencing

Effects of miRNA-23a-3p overexpression on cell proliferation in HG-treated H9c2 cells. ( a ) RT-qPCR to detect the expression level of miRNA-23a-3p in HG-treated H9c2 cells (**** p < 0.0001 vs. NG group). ( b ) H9c2 cells were exposed to glucose at concentrations of 5 mM (NG), 35 mM (HG), and 5 mM glucose + 30 mM mannitol (NG + Mannitol) for 24, 48, and 72 h (**** p < 0.0001 vs. NG group). Cell viability was assessed by a CCK-8 assay. ( c ) H9c2 cell viability was significantly increased following treatment with the miRNA-23a-3p mimics (** p < 0.01 vs. NC-mimics). ( d ) Representative images illustrating the EdU and Hoechst staining of H9c2 cells exposed to 5 mM and 35 mM glucose and transfected with NC mimics or miRNA-23a-3p mimics. ( e ) Analysis of EdU-positive cardiomyocytes by an EdU incorporation assay. The percentages of proliferative H9c2 cells were calculated ( n > 500). Compared with the NC-mimics group, the miRNA-23a-3p-mimics group had increased EdU incorporation (** p < 0.01). CCK-8 Cell Counting Kit-8, EdU 5-ethynyl-2′-deoxyuridine, NG normal glucose, HG high glucose, NC negative control.

Journal: In Vitro Cellular & Developmental Biology. Animal

Article Title: Upregulation of miRNA-23a-3p rescues high glucose-induced cell apoptosis and proliferation inhibition in cardiomyocytes

doi: 10.1007/s11626-020-00518-6

Figure Lengend Snippet: Effects of miRNA-23a-3p overexpression on cell proliferation in HG-treated H9c2 cells. ( a ) RT-qPCR to detect the expression level of miRNA-23a-3p in HG-treated H9c2 cells (**** p < 0.0001 vs. NG group). ( b ) H9c2 cells were exposed to glucose at concentrations of 5 mM (NG), 35 mM (HG), and 5 mM glucose + 30 mM mannitol (NG + Mannitol) for 24, 48, and 72 h (**** p < 0.0001 vs. NG group). Cell viability was assessed by a CCK-8 assay. ( c ) H9c2 cell viability was significantly increased following treatment with the miRNA-23a-3p mimics (** p < 0.01 vs. NC-mimics). ( d ) Representative images illustrating the EdU and Hoechst staining of H9c2 cells exposed to 5 mM and 35 mM glucose and transfected with NC mimics or miRNA-23a-3p mimics. ( e ) Analysis of EdU-positive cardiomyocytes by an EdU incorporation assay. The percentages of proliferative H9c2 cells were calculated ( n > 500). Compared with the NC-mimics group, the miRNA-23a-3p-mimics group had increased EdU incorporation (** p < 0.01). CCK-8 Cell Counting Kit-8, EdU 5-ethynyl-2′-deoxyuridine, NG normal glucose, HG high glucose, NC negative control.

Article Snippet: Real-time qPCR was performed with SuperReal PreMix Plus (FP 205, TianGen) and the miRcute Plus miRNA qPCR Detection Kit (FP 411, TianGen) on an Exicycler 96 PCR thermal block (Light Cycler 480, Roche, Basel, Switzerland).

Techniques: Over Expression, Quantitative RT-PCR, Expressing, CCK-8 Assay, Staining, Transfection, Cell Counting, Negative Control

Data analysis of the cell cycle duration. Manual tracking of 200 cell cycle trajectories was performed (20-min frame interval). For example, a , b , c , and d show a single-cell trajectory evaluated with time-lapse acquisition exhibiting one cell division. ( a ) The measured cell cycle length is 14.33 h in a cell in the NG group. ( b ) The measured cell cycle length is 17 h in a cell in the HG group. ( c ) The measured cell cycle length is 15.33 h in a cell in the NC-mimics group. ( d ) The measured cell cycle length is 12.33 h in a cell in the miRNA-23a-3p-mimics group. Each cropped image is 80 × 80 μm 2 . The beginning of cell division is denoted by white circles. ( e ) Fifty cells per group were randomly tracked to calculate the cell cycle duration by detecting the occurrence of cell division in the cell track ( n = 50). The statistical analysis of the cell cycle duration in the different groups was performed using one-way ANOVA. ns not significant, ** p < 0.01, *** p < 0.001.

Journal: In Vitro Cellular & Developmental Biology. Animal

Article Title: Upregulation of miRNA-23a-3p rescues high glucose-induced cell apoptosis and proliferation inhibition in cardiomyocytes

doi: 10.1007/s11626-020-00518-6

Figure Lengend Snippet: Data analysis of the cell cycle duration. Manual tracking of 200 cell cycle trajectories was performed (20-min frame interval). For example, a , b , c , and d show a single-cell trajectory evaluated with time-lapse acquisition exhibiting one cell division. ( a ) The measured cell cycle length is 14.33 h in a cell in the NG group. ( b ) The measured cell cycle length is 17 h in a cell in the HG group. ( c ) The measured cell cycle length is 15.33 h in a cell in the NC-mimics group. ( d ) The measured cell cycle length is 12.33 h in a cell in the miRNA-23a-3p-mimics group. Each cropped image is 80 × 80 μm 2 . The beginning of cell division is denoted by white circles. ( e ) Fifty cells per group were randomly tracked to calculate the cell cycle duration by detecting the occurrence of cell division in the cell track ( n = 50). The statistical analysis of the cell cycle duration in the different groups was performed using one-way ANOVA. ns not significant, ** p < 0.01, *** p < 0.001.

Article Snippet: Real-time qPCR was performed with SuperReal PreMix Plus (FP 205, TianGen) and the miRcute Plus miRNA qPCR Detection Kit (FP 411, TianGen) on an Exicycler 96 PCR thermal block (Light Cycler 480, Roche, Basel, Switzerland).

Techniques:

Effects of miRNA-23a-3p overexpression on cell apoptosis in HG-treated H9c2 cells apoptosis analysis by flow cytometry. ( a ) H9c2 cells treated with NG, NG + mannitol (30 mM) and HG. The rates of early cardiomyocyte apoptosis were significantly increased in the HG group (*** p < 0.001 vs. NG group). ( b ) H9c2 cells exposed to HG and transfected with NC or miRNA-23a-3p mimics. The rates of early cardiomyocyte apoptosis were significantly decreased in the miRNA-23a-3p-mimics group (**** p < 0.0001 vs. NC mimics). FITC fluorescein isothiocyanate, PI propidium iodide, Q2 late apoptotic rates, Q3 early apoptotic rates.

Journal: In Vitro Cellular & Developmental Biology. Animal

Article Title: Upregulation of miRNA-23a-3p rescues high glucose-induced cell apoptosis and proliferation inhibition in cardiomyocytes

doi: 10.1007/s11626-020-00518-6

Figure Lengend Snippet: Effects of miRNA-23a-3p overexpression on cell apoptosis in HG-treated H9c2 cells apoptosis analysis by flow cytometry. ( a ) H9c2 cells treated with NG, NG + mannitol (30 mM) and HG. The rates of early cardiomyocyte apoptosis were significantly increased in the HG group (*** p < 0.001 vs. NG group). ( b ) H9c2 cells exposed to HG and transfected with NC or miRNA-23a-3p mimics. The rates of early cardiomyocyte apoptosis were significantly decreased in the miRNA-23a-3p-mimics group (**** p < 0.0001 vs. NC mimics). FITC fluorescein isothiocyanate, PI propidium iodide, Q2 late apoptotic rates, Q3 early apoptotic rates.

Article Snippet: Real-time qPCR was performed with SuperReal PreMix Plus (FP 205, TianGen) and the miRcute Plus miRNA qPCR Detection Kit (FP 411, TianGen) on an Exicycler 96 PCR thermal block (Light Cycler 480, Roche, Basel, Switzerland).

Techniques: Over Expression, Flow Cytometry, Transfection

Expression of cell proliferation- and apoptosis-related factors in H9c2 cells under HG conditions and in HG-treated H9c2 cells transfected with miRNA-23a-3p mimics. The mRNA expression levels of Ccna2 , Ccne1 , Ccnd1 , Ccnd2 , Ccnb1 , and Bcl-2 were determined by qRT-PCR. All data are expressed as the mean ± standard error of mean. ns not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (HG vs. NG, NC mimics vs. HG or miRNA-23a-3p vs. NC mimics).

Journal: In Vitro Cellular & Developmental Biology. Animal

Article Title: Upregulation of miRNA-23a-3p rescues high glucose-induced cell apoptosis and proliferation inhibition in cardiomyocytes

doi: 10.1007/s11626-020-00518-6

Figure Lengend Snippet: Expression of cell proliferation- and apoptosis-related factors in H9c2 cells under HG conditions and in HG-treated H9c2 cells transfected with miRNA-23a-3p mimics. The mRNA expression levels of Ccna2 , Ccne1 , Ccnd1 , Ccnd2 , Ccnb1 , and Bcl-2 were determined by qRT-PCR. All data are expressed as the mean ± standard error of mean. ns not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (HG vs. NG, NC mimics vs. HG or miRNA-23a-3p vs. NC mimics).

Article Snippet: Real-time qPCR was performed with SuperReal PreMix Plus (FP 205, TianGen) and the miRcute Plus miRNA qPCR Detection Kit (FP 411, TianGen) on an Exicycler 96 PCR thermal block (Light Cycler 480, Roche, Basel, Switzerland).

Techniques: Expressing, Transfection, Quantitative RT-PCR

The rescue effects of different doses of miRNA-23a-3p on HG-induced cell proliferation HG-treated H9c2 cells transfected with NC mimics and different doses of miRNA-23a-3p mimics (3, 6, 9, 12, 18, 24, and 30 pmol/well). Cell viability was determined by a CCK-8 assay. ns not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. NC mimics.

Journal: In Vitro Cellular & Developmental Biology. Animal

Article Title: Upregulation of miRNA-23a-3p rescues high glucose-induced cell apoptosis and proliferation inhibition in cardiomyocytes

doi: 10.1007/s11626-020-00518-6

Figure Lengend Snippet: The rescue effects of different doses of miRNA-23a-3p on HG-induced cell proliferation HG-treated H9c2 cells transfected with NC mimics and different doses of miRNA-23a-3p mimics (3, 6, 9, 12, 18, 24, and 30 pmol/well). Cell viability was determined by a CCK-8 assay. ns not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. NC mimics.

Article Snippet: Real-time qPCR was performed with SuperReal PreMix Plus (FP 205, TianGen) and the miRcute Plus miRNA qPCR Detection Kit (FP 411, TianGen) on an Exicycler 96 PCR thermal block (Light Cycler 480, Roche, Basel, Switzerland).

Techniques: Transfection, CCK-8 Assay

The repair ability of miRNA-23a-3p in HG-treated H9c2 cells depends on the manipulation time H9c2 cells were transfected with miRNA-23a-3p 6 h before HG treatment (group 1), at the same time (group 2), or after 6 h of HG treatment (group 3). Cell viability was determined by a CCK-8 assay. ** p < 0.01, group 1 vs. group 2; *** p < 0.001, group 2 vs. group 3; **** p < 0.0001, group 1 vs. group 3.

Journal: In Vitro Cellular & Developmental Biology. Animal

Article Title: Upregulation of miRNA-23a-3p rescues high glucose-induced cell apoptosis and proliferation inhibition in cardiomyocytes

doi: 10.1007/s11626-020-00518-6

Figure Lengend Snippet: The repair ability of miRNA-23a-3p in HG-treated H9c2 cells depends on the manipulation time H9c2 cells were transfected with miRNA-23a-3p 6 h before HG treatment (group 1), at the same time (group 2), or after 6 h of HG treatment (group 3). Cell viability was determined by a CCK-8 assay. ** p < 0.01, group 1 vs. group 2; *** p < 0.001, group 2 vs. group 3; **** p < 0.0001, group 1 vs. group 3.

Article Snippet: Real-time qPCR was performed with SuperReal PreMix Plus (FP 205, TianGen) and the miRcute Plus miRNA qPCR Detection Kit (FP 411, TianGen) on an Exicycler 96 PCR thermal block (Light Cycler 480, Roche, Basel, Switzerland).

Techniques: Transfection, CCK-8 Assay