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

Figure 4.

Figure 4

Figure 4

YSV inhibited the expression and activity of FAK in human cancer lung cells.

Notes: (A) Real-time PCR analysis of FAK mRNA level 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 FAK mRNA level. The products were quantified using β-actin as an internal reference. (B) Western blot for FAK in the 95D, A549, and NCI-H1299 cells after treated with YSV in vitro. Total proteins of 95D, A549, and NCI-H1299 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 antibody against target protein. The products are reported as the target protein/β-actin densitometric ratio calculated by the TotalLab software to compute the relative expression of proteins. (C) Western blot for phosphorylation of FAK Tyr397 and Tyr576/577 in the 95D, A549, and NCI-H1299 cells after treated with YSV in vitro. Total proteins of 95D, A549, and NCI-H1299 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 antibody against target protein. 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; FAK, focal adhesion kinase; PCR, polymerase chain reaction; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; PVDF, polyvinylidene difluoride; ANOVA, analysis of variance.