Journal: Experimental and Therapeutic Medicine

Article Title: hsa-miR-15b-5p regulates the proliferation and apoptosis of human vascular smooth muscle cells by targeting the ACSS2/PTGS2 axis

doi: 10.3892/etm.2021.10642

Figure Lengend Snippet: Expression levels of miR-15b-5p in human aortic vascular smooth muscle cells after different transfections, as determined by reverse transcription quantitative-PCR. Results are presented as the average+standard deviation of three independent experiments. * P<0.05 vs. mimic NC; # P<0.05 vs. miR-15b-5p mimics; & P<0.05 vs. inhibitor NC. miR, microRNA; NC, negative control.

Article Snippet: Empty pcDNA3.1 vector, ACSS2 overexpression plasmid (pcDNA3.1/ACSS2), miR-15b mimics/inhibitors and mimic/inhibitor negative controls (NC) were obtained from RiboBio Co., Ltd.. At 24 h before the transient transfections, an appropriate number of human aortic VSMCs were seeded into six-well plates with 2 ml of SMCM and grown to 60-70% confluency at 37 ̊C in an atmosphere containing 5% CO 2 .

Techniques: Expressing, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Standard Deviation, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: hsa-miR-15b-5p regulates the proliferation and apoptosis of human vascular smooth muscle cells by targeting the ACSS2/PTGS2 axis

doi: 10.3892/etm.2021.10642

Figure Lengend Snippet: Proliferation of human aortic VSMCs after different transfections, as determined by CCK-8 assay. (A) miR-15b-5p mimics and NC groups. (B) miR-15b-5p inhibitors and NC groups. (C) Rescue experiments by simultaneous overexpression of ACSS2 in aortic VSMCs transfected with miR-15b-5p mimics. The OD was detected at 0, 24, 48 and 72 h after transfection. Results are presented as the average ± standard deviation of four independent experiments. * P<0.05 vs. mimic NC; & P<0.05 vs. inhibitor NC; $ P<0.05 vs. miR-15b-5p mimics+pcDNA3.1 empty vector. VSMCs, vascular smooth muscle cells; miR, microRNA; NC, negative control; ACSS2, acyl-CoA synthetase short chain family member 2; OD, optical density.

Article Snippet: Empty pcDNA3.1 vector, ACSS2 overexpression plasmid (pcDNA3.1/ACSS2), miR-15b mimics/inhibitors and mimic/inhibitor negative controls (NC) were obtained from RiboBio Co., Ltd.. At 24 h before the transient transfections, an appropriate number of human aortic VSMCs were seeded into six-well plates with 2 ml of SMCM and grown to 60-70% confluency at 37 ̊C in an atmosphere containing 5% CO 2 .

Techniques: Transfection, CCK-8 Assay, Over Expression, Standard Deviation, Plasmid Preparation, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: hsa-miR-15b-5p regulates the proliferation and apoptosis of human vascular smooth muscle cells by targeting the ACSS2/PTGS2 axis

doi: 10.3892/etm.2021.10642

Figure Lengend Snippet: Apoptosis of human aortic VSMCs at 48 h after different transfections, as determined by flow cytometry assay. (A) miR-15b-5p mimics and NC groups. (B) miR-15b-5p inhibitors and NC groups. (C) Rescue experiments by simultaneous overexpression of ACSS2 in aortic VSMCs transfected with miR-15b-5p mimics. Representative flow cytometry plots and quantifications are shown. Results are presented as the average+standard deviation of two independent experiments. * P<0.05 vs. mimic NC; & P<0.05 vs. inhibitor NC; $ P<0.05 vs. miR-15b-5p mimics+pcDNA3.1 empty vector. VSMCs, vascular smooth muscle cells; miR, microRNA; NC, negative control; ACSS2, acyl-CoA synthetase short chain family member 2.

Article Snippet: Empty pcDNA3.1 vector, ACSS2 overexpression plasmid (pcDNA3.1/ACSS2), miR-15b mimics/inhibitors and mimic/inhibitor negative controls (NC) were obtained from RiboBio Co., Ltd.. At 24 h before the transient transfections, an appropriate number of human aortic VSMCs were seeded into six-well plates with 2 ml of SMCM and grown to 60-70% confluency at 37 ̊C in an atmosphere containing 5% CO 2 .

Techniques: Transfection, Flow Cytometry, Over Expression, Standard Deviation, Plasmid Preparation, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: hsa-miR-15b-5p regulates the proliferation and apoptosis of human vascular smooth muscle cells by targeting the ACSS2/PTGS2 axis

doi: 10.3892/etm.2021.10642

Figure Lengend Snippet: ACSS2 is a direct target of miR-15b-5p. (A) Schematic of the predicted binding site between miR-15b-5p and the 3'-UTR of ACSS2. (B) Luciferase reporter assays were conducted to confirm the binding ability between miR-15b-5p and ACSS2. WT or MUT constructs of the 3'-UTR of ACSS2 were co-transfected with miR-15b-5p mimics or NC in HEK-293T cells. The firefly luciferase activity was normalized against Renilla luciferase activity. Results are presented as the average+standard deviation of four independent experiments. * P<0.05 vs. mimic NC; # P<0.05 vs. miR-15b-5p mimics. ACSS2, acyl-CoA synthetase short chain family member 2; miR, microRNA; UTR, untranslated region; WT, wild-type; MUT; mutant; NC, negative control.

Article Snippet: Empty pcDNA3.1 vector, ACSS2 overexpression plasmid (pcDNA3.1/ACSS2), miR-15b mimics/inhibitors and mimic/inhibitor negative controls (NC) were obtained from RiboBio Co., Ltd.. At 24 h before the transient transfections, an appropriate number of human aortic VSMCs were seeded into six-well plates with 2 ml of SMCM and grown to 60-70% confluency at 37 ̊C in an atmosphere containing 5% CO 2 .

Techniques: Binding Assay, Luciferase, Construct, Transfection, Activity Assay, Standard Deviation, Mutagenesis, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: hsa-miR-15b-5p regulates the proliferation and apoptosis of human vascular smooth muscle cells by targeting the ACSS2/PTGS2 axis

doi: 10.3892/etm.2021.10642

Figure Lengend Snippet: ACSS2 and PTGS2 expression levels are regulated by miR-15b-5p. (A) RT-qPCR was performed to confirm the successful overexpression of ACSS2. (B) RT-qPCR was performed to measure the mRNA expression levels of ACSS2 after miR-15b-5p transfection treatments. (C) Representative images from western blot analysis showing the protein expression levels of ACSS2 and PTGS2 in the different transfection groups. (D) Quantitative results of ACSS2 protein. (E) Quantitative results of PTGS2 protein. Results are presented as the average+standard deviation of three independent experiments for mRNAs and two independent experiments for proteins. £ P<0.05 vs. pcDNA3.1; * P<0.05 vs. mimic NC; # P<0.05 vs. miR-15b-5p mimics; & P<0.05 vs. inhibitor NC; @ P<0.05 vs. miR-15b-5p inhibitors; $ P<0.05 vs. miR-15b-5p mimics+pcDNA3.1 empty vector. ACSS2, acyl-CoA synthetase short chain family member 2; PTGS2, prostaglandin-endoperoxide synthase 2; miR, microRNA; RT-qPCR, reverse transcription-quantitative PCR; NC, negative control.

Article Snippet: Empty pcDNA3.1 vector, ACSS2 overexpression plasmid (pcDNA3.1/ACSS2), miR-15b mimics/inhibitors and mimic/inhibitor negative controls (NC) were obtained from RiboBio Co., Ltd.. At 24 h before the transient transfections, an appropriate number of human aortic VSMCs were seeded into six-well plates with 2 ml of SMCM and grown to 60-70% confluency at 37 ̊C in an atmosphere containing 5% CO 2 .

Techniques: Expressing, Quantitative RT-PCR, Over Expression, Transfection, Western Blot, Standard Deviation, Plasmid Preparation, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control