Journal: PLoS ONE

Article Title: 14-3-3β Promotes Migration and Invasion of Human Hepatocellular Carcinoma Cells by Modulating Expression of MMP2 and MMP9 through PI3K/Akt/NF-κB Pathway

doi: 10.1371/journal.pone.0146070

Figure Lengend Snippet: (A) Matrigel-free Transwell assay was conducted to assess the migration ability of SMCC-7721 and Hep3B cells that were transfected with 14-3-3β expression construct or the empty vector pcDNA3.1. (B) Matrigel-based Transwell assay was performed to examine the invasiveness of SMCC-7721 and Hep3B with exogenous expression of 14-3-3β. (C) Matrigel-free migration assay and (D) Matrigel-based invasion assay were performed in CSQT-2 cells with and without expression of 14-3-3β-specific siRNA. This figure shows the representative images and the statistical analysis of three independent experiments. Values are expressed as the mean ± standard deviation. *** p <0.001.

Article Snippet: Cell migration ability was assessed by the Transwell assay with uncoated upper chambers (Corning, New York, USA) [ ], whereas cell invasion was examined by the Transwell assay with the upper chamber pre-coated with phenol red-free Matrigel (BD Biosciences, San Jose, CA, USA).

Techniques: Transwell Assay, Migration, Transfection, Expressing, Construct, Plasmid Preparation, Invasion Assay, Standard Deviation

Journal: PLoS ONE

Article Title: 14-3-3β Promotes Migration and Invasion of Human Hepatocellular Carcinoma Cells by Modulating Expression of MMP2 and MMP9 through PI3K/Akt/NF-κB Pathway

doi: 10.1371/journal.pone.0146070

Figure Lengend Snippet: (A) Expression of 14-3-3β, MMP2 and MMP9 in 14-3-3β-transfected SMCC-7721 and Hep3B cells. (B) Western blot analysis of phosphorylation status of various signaling proteins and the level of nuclear p65 in 14-3-3β-overexpressing HCC cells. β-actin was used as the internal control for total or cytosolic proteins, while Lamin A served as the internal control for nuclear proteins. The 14-3-3β-transfected Hep3B cells were treated with (C) LY294002 (20 μM), the inhibitor of PI3K/Akt signaling, or (D) PDTC (10 μM), the inhibitor of NF-κB signaling, for 5 h, followed by western blot analysis of the expression of MMP2 and MMP9. (E) The invasiveness of 14-3-3β-overexpressing cells with and without LY294002 (20 μM) or PDTC (10 μM) treatment was assessed by Matrigel-based Transwell assay. Values are expressed as the mean ± standard deviation of three independent experiments. *** p <0.001. (F) Western blot analysis was performed to determine the activation status of p-Akt and NF-κB in 14-3-3β-overexpressing cells after treating the cells with LY294002 (20 μM) for 5 h.

Article Snippet: Cell migration ability was assessed by the Transwell assay with uncoated upper chambers (Corning, New York, USA) [ ], whereas cell invasion was examined by the Transwell assay with the upper chamber pre-coated with phenol red-free Matrigel (BD Biosciences, San Jose, CA, USA).

Techniques: Expressing, Transfection, Western Blot, Transwell Assay, Standard Deviation, Activation Assay