Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2016 Mar 3;10:649–663. doi: 10.2147/DDDT.S86284

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2016 Huang et al. This work is published and licensed by Dove Medical Press Limited

The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed.

PMC Copyright notice

YSV inhibited the expression and activity of MMP-2 and MMP-9.

Notes: (A) Gelatin zymography assay for MMP-2 and MMP-9 activities from supernatant of 95D, A549, and NCI-H1299 cells after treated with YSV in vitro. Supernatants of lung cancer cells treated with different concentrations of YSV for 48 hours were collected and loaded onto 10% acrylamide (containing 0.1% gelatin) gels. After electrophoresis, the gels were washed, incubated, stained, and destained. Gelatinolytic activities were identified as clear bands on a background. The relative gelatinolytic activities of MMP-2 and MMP-9 were quantified densitometrically in relation to the activity without YSV. (B) Real-time PCR analysis of MMP-2 and MMP-9 mRNA levels in the 95D, A549, and NCI-H1299 cells after treated with YSV in vitro. Real-time PCR was performed on total RNA for detection of mRNA level of MMP-2 and MMP-9. The products were quantified using β-actin as an internal reference. (C) 2% agarose gel electrophoresis of real-time PCR production. (D) Western blot analysis of the levels of MMP-2 and MMP-9 in the 95D, A549, and NCI-H1299 cells after treated with YSV in vitro. Total proteins of lung cancer cells treated with different concentrations of YSV for 48 hours were extracted and separated by SDS-PAGE. After electrophoresis, the protein was transferred to PVDF membrane, and detection was performed with antibodies against target proteins. The products are reported as the target protein/β-actin densitometric ratio calculated by the TotalLab software to compute the relative expression of proteins. ^*P<0.05, compared to control group, tested using one-way ANOVA and Student–Newman–Keuls test (N=3).

Abbreviations: YSV, tyroservatide; MMP, matrix metalloproteinase; PVDF, polyvinylidene difluoride; PCR, polymerase chain reaction; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; ANOVA, analysis of variance.

YSV inhibited the expression and activity of MMP-2 and MMP-9.

Notes: (A) Gelatin zymography assay for MMP-2 and MMP-9 activities from supernatant of 95D, A549, and NCI-H1299 cells after treated with YSV in vitro. Supernatants of lung cancer cells treated with different concentrations of YSV for 48 hours were collected and loaded onto 10% acrylamide (containing 0.1% gelatin) gels. After electrophoresis, the gels were washed, incubated, stained, and destained. Gelatinolytic activities were identified as clear bands on a background. The relative gelatinolytic activities of MMP-2 and MMP-9 were quantified densitometrically in relation to the activity without YSV. (B) Real-time PCR analysis of MMP-2 and MMP-9 mRNA levels in the 95D, A549, and NCI-H1299 cells after treated with YSV in vitro. Real-time PCR was performed on total RNA for detection of mRNA level of MMP-2 and MMP-9. The products were quantified using β-actin as an internal reference. (C) 2% agarose gel electrophoresis of real-time PCR production. (D) Western blot analysis of the levels of MMP-2 and MMP-9 in the 95D, A549, and NCI-H1299 cells after treated with YSV in vitro. Total proteins of lung cancer cells treated with different concentrations of YSV for 48 hours were extracted and separated by SDS-PAGE. After electrophoresis, the protein was transferred to PVDF membrane, and detection was performed with antibodies against target proteins. The products are reported as the target protein/β-actin densitometric ratio calculated by the TotalLab software to compute the relative expression of proteins. ^*P<0.05, compared to control group, tested using one-way ANOVA and Student–Newman–Keuls test (N=3).

Abbreviations: YSV, tyroservatide; MMP, matrix metalloproteinase; PVDF, polyvinylidene difluoride; PCR, polymerase chain reaction; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; ANOVA, analysis of variance.