Journal: Experimental and Therapeutic Medicine

Article Title: COL6A1 knockdown suppresses cell proliferation and migration in human aortic vascular smooth muscle cells

doi: 10.3892/etm.2019.7798

Figure Lengend Snippet: COL6A1 knockdown attenuated cell viability in PDGF-BB-stimulated HA-VSMCs. (A) Expression levels of COL6A1 mRNA were determined using reverse transcription-quantitative PCR following si-COL6A1 transfection. (B) COL6A1 protein levels were determined using western blotting after si-COL6A1 transfection. (C) Quantified densitometry results from the western blot analysis. (D) Cell viability after 12, 24 and 48 h were measured using Cell Counting Kit-8 assay after si-COL6A1 transfection and PDGF-BB stimulation. Each experiment was repeated five times. *P<0.05 and **P<0.01. HA-VSMC, human aortic-vascular smooth muscle cells; COL6A1, collagen type VI α1 chain; PDGF-BB, platelet-derived growth factor; si, short interfering; Cntl, control; EV, siRNA-control.

Article Snippet: Cntl, EV and si-COL6A1 cells (5×10 5 cells) treated or untreated with 20 ng/ml PDGF-BB were lysed using RIPA lysis buffer (Boster Biological Technology) for 20 min, with the proteins quantified using Bicinchoninic Acid protein assay kit (Pierce; Thermo Fisher Scientific, Inc.).

Techniques: Knockdown, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Transfection, Western Blot, Cell Counting, Derivative Assay, Control

Journal: Experimental and Therapeutic Medicine

Article Title: COL6A1 knockdown suppresses cell proliferation and migration in human aortic vascular smooth muscle cells

doi: 10.3892/etm.2019.7798

Figure Lengend Snippet: COL6A1 interference inhibited PDGF-BB-stimulated HA-VSMC migration. (A) Wound closure was measured in HA-VSMCs transfected with si-COL6A1 following PDGF-BB stimulation using wound healing assay. (B) Relative wound closure for each condition was quantified. Each experiment was repeated five times. **P<0.01. HA-VSMC, human aortic-vascular smooth muscle cells; COL6A1, collagen type VI α1 chain; PDGF-BB, platelet-derived growth factor; si, short interfering; Cntl, control; EV, siRNA-control.

Article Snippet: Cntl, EV and si-COL6A1 cells (5×10 5 cells) treated or untreated with 20 ng/ml PDGF-BB were lysed using RIPA lysis buffer (Boster Biological Technology) for 20 min, with the proteins quantified using Bicinchoninic Acid protein assay kit (Pierce; Thermo Fisher Scientific, Inc.).

Techniques: Migration, Transfection, Wound Healing Assay, Derivative Assay, Control

Journal: Experimental and Therapeutic Medicine

Article Title: COL6A1 knockdown suppresses cell proliferation and migration in human aortic vascular smooth muscle cells

doi: 10.3892/etm.2019.7798

Figure Lengend Snippet: COL6A1 knockdown inhibited HA-VSMC invasion following PDGF-BB stimulation. (A) HA-VSMC invasion was measured using Matrigel-coated Transwell assay after si-COL6A1 transfection and PDGF-BB stimulation. (B) Relative invasion for each condition were quantified. Each experiment was repeated five times. **P<0.01. HA-VSMC, human aortic-vascular smooth muscle cells; COL6A1, collagen type VI α1 chain; PDGF-BB, platelet-derived growth factor; si, short interfering; Cntl, control; EV, siRNA-control.

Article Snippet: Cntl, EV and si-COL6A1 cells (5×10 5 cells) treated or untreated with 20 ng/ml PDGF-BB were lysed using RIPA lysis buffer (Boster Biological Technology) for 20 min, with the proteins quantified using Bicinchoninic Acid protein assay kit (Pierce; Thermo Fisher Scientific, Inc.).

Techniques: Knockdown, Transwell Assay, Transfection, Derivative Assay, Control

Journal: Experimental and Therapeutic Medicine

Article Title: COL6A1 knockdown suppresses cell proliferation and migration in human aortic vascular smooth muscle cells

doi: 10.3892/etm.2019.7798

Figure Lengend Snippet: COL6A1 regulate the expression of factors associated with migration and invasion in HA-VSMCs following PDGF-BB stimulation. (A) Reverse transcription-quantitative PCR was performed to determine the levels of mRNA expression of FN, (B) MMP-2, (C) MMP-9 and (D) TIMP-2 after si-COL6A1 transfection and PDGF-BB stimulation. (E-I) Western blot analysis was performed to determine the protein expression levels of (F) FN, (G) MMP-2, (H) MMP-9 and (I) TIMP-2 after si-COL6A1 transfection and PDGF-BB stimulation. Each experiment was repeated five times. *P<0.05, and **P<0.01. HA-VSMC, human aortic-vascular smooth muscle cells; COL6A1, collagen type VI α1 chain; PDGF-BB, platelet-derived growth factor; FN, fibronectin; MMP, matrix metalloproteinase; TIMP-2, tissue inhibitor of metalloproteinase-2; si, short interfering; Cntl, control; EV, siRNA-control.

Article Snippet: Cntl, EV and si-COL6A1 cells (5×10 5 cells) treated or untreated with 20 ng/ml PDGF-BB were lysed using RIPA lysis buffer (Boster Biological Technology) for 20 min, with the proteins quantified using Bicinchoninic Acid protein assay kit (Pierce; Thermo Fisher Scientific, Inc.).

Techniques: Expressing, Migration, Reverse Transcription, Real-time Polymerase Chain Reaction, Transfection, Western Blot, Derivative Assay, Control

Journal: Experimental and Therapeutic Medicine

Article Title: COL6A1 knockdown suppresses cell proliferation and migration in human aortic vascular smooth muscle cells

doi: 10.3892/etm.2019.7798

Figure Lengend Snippet: AKT-mTOR pathway was inhibited by COL6A1 knockdown in HA-VSMCs following PDGF-BB stimulation. (A-C) The levels of phosphorylation relative to the total expression of their respective proteins of (B) AKT and (C) mTOR in HA-VSMCs were analyzed and quantified using western blot analysis following si-COL6A1 transfection and PDGF-BB stimulation. Each experiment was repeated five times. *P<0.05 and **P<0.01. HA-VSMC, human aortic-vascular smooth muscle cells; COL6A1, collagen type VI α1 chain; PDGF-BB, platelet-derived growth factor; si, short interfering; Cntl, control; EV, siRNA-control.

Article Snippet: Cntl, EV and si-COL6A1 cells (5×10 5 cells) treated or untreated with 20 ng/ml PDGF-BB were lysed using RIPA lysis buffer (Boster Biological Technology) for 20 min, with the proteins quantified using Bicinchoninic Acid protein assay kit (Pierce; Thermo Fisher Scientific, Inc.).

Techniques: Knockdown, Phospho-proteomics, Expressing, Western Blot, Transfection, Derivative Assay, Control