si fak  (Santa Cruz Biotechnology)

Journal: PLoS ONE

Article Title: Bcl-w Enhances Mesenchymal Changes and Invasiveness of Glioblastoma Cells by Inducing Nuclear Accumulation of β-Catenin

doi: 10.1371/journal.pone.0068030

Figure Lengend Snippet: A , the indicated U251 cell transfectants were incubated in serum-free medium in the presence or absence of PI3K inhibitor (LY294002 (LY); 10 µmol/L) or Akt inhibitor (Akt-I; 10 µmol/L) for 1 hour. Expression levels and activities of p-Akt, p-GSK3β, β-catenin, TCF-4 and MMP-2 proteins were compared using Western blotting. B , cells treated with PI3K inhibitor or Akt inhibitor in the lower compartments of the invasion chambers for 24 hours, respectively. Invasive potential of treated cells was compared. *, p< 0.05 versus untreated control, n = 5. C , β-catenin and TCF-4 siRNAs (20nM) were introduced into vector or Bcl-w overexpressing cells, and cellular levels of β-catenin, TCF-4, MMP-2 and p-FAK compared after 24 hours of incubation using Western blotting with β-actin as a loading control. D , invasive potential of the indicated transfectants was compared. *, p< 0.05, n = 5.

Article Snippet: The following small interfere RNAs purchased from; silencer negative control siRNA, si-Bcl-w-1 and si-Bcl-w-2 (Ambion, Cambridge, MA); si-vimentin, si-Twist1, si-Snail si-β-catenin, si-TCF-4, si-MMP-2 and si-FAK (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Incubation, Expressing, Western Blot, Plasmid Preparation

Journal: PLoS ONE

Article Title: Bcl-w Enhances Mesenchymal Changes and Invasiveness of Glioblastoma Cells by Inducing Nuclear Accumulation of β-Catenin

doi: 10.1371/journal.pone.0068030

Figure Lengend Snippet: A , right image, MMP-2 siRNA (20nM) was introduced into the indicated U251 transfectants, and after 24 hours of incubation, p-FAK (Y397) and MMP-2 protein levels were compared using Western blotting. Left image, invasion assays were performed using small interfering RNA MMP-2-treated and untreated cells. *, p< 0.01 versus untreated control, n = 5. B , top image, FAK siRNA was introduced into the indicated transfectants, and after 24 hours of incubation, cellular levels of FAK, p-FAK and MMP-2 compared using Western blotting. Bottom plots, invasion assays were conducted using the indicated cells. *, p< 0.05, n = 5. C , top images, vector- and Bcl-w-expressing cells were transiently transfected with expression vectors for HA-tagged dominant-negative FAK mutant (FAKY397F). After 24 hours of incubation, expression of the introduced mutants in cells was verified by Western blotting. Bottom plots, invasive potentials of the indicated cells were compared. *, p < 0.05, n = 5.

Article Snippet: The following small interfere RNAs purchased from; silencer negative control siRNA, si-Bcl-w-1 and si-Bcl-w-2 (Ambion, Cambridge, MA); si-vimentin, si-Twist1, si-Snail si-β-catenin, si-TCF-4, si-MMP-2 and si-FAK (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Incubation, Western Blot, Small Interfering RNA, Plasmid Preparation, Expressing, Transfection, Dominant Negative Mutation, Mutagenesis