Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Inhibition of cell-matrix adhesions prevents cartilage chondrocyte death following impact injury.

doi: 10.1002/jor.22523

Figure Lengend Snippet: Figure 3. Confocal microscopy and cell viability immediately after impact injury. (A) Confocal micrographs show live (green) and dead (red) chondrocytes in an impact site in an un-treated control explant, and in explants treated with 10 mM SFKi and either 10 or 100 mM FAKi. Compared to control, fewer dead chondrocytes were observed in SFKs or FAKi treated groups. (B) Statistical analysis revealed that chondrocyte viability was significantly higher in SFKs or FAKi treated explants compared to control. Between two tested concentrations, 100 mM FAKi was more effective than 10 mM. Asterisk represents statistically significant (p < 0.05, p < 0.01). Bars ¼ 500 mm.

Article Snippet: Published by Wiley Periodicals, Inc. 448 JOURNAL OF ORTHOPAEDIC RESEARCH MARCH 2014 After 2 days, the explants were randomly distributed and were treated with fresh culture medium containing 10 or 100mM focal adhesion kinase inhibitor (FAKi) (Santa Cruz Biotechnology, Dallas, TX) to block phosphorylation of FAK at the kinase domain (Try 397) or were treated with fresh culture medium containing 10mM Src family kinase inhibitor (SFKi) (Selleckchem, Houston, TX) to block phosphorylation of SFKs at kinase domain (Tyr 416) for 2h.

Techniques: Confocal Microscopy, Control

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Inhibition of cell-matrix adhesions prevents cartilage chondrocyte death following impact injury.

doi: 10.1002/jor.22523

Figure Lengend Snippet: Figure 5. Kinetics of FAK and SFKi by western blot analysis. (A) Immunoblot analysis showed that treatment with 10 ng/ml IL-1b and 100 ng/ml TNF-a for 30 min significantly increased SFKs phosphorylation at Tyr 416. 0.1, 1, and 10 mM SFKi diminished this response dose dependently. (B) Analysis of the integrated densities of the bands with the phosphor- to total SFKs ratio. C represents untreated control and Cþ represents 10 ng/ml IL-1b and 100 ng/ml TNF-a treated only. (C) Immunoblots show that treatment with 100 nM fMLF for 30 min did not enhance FAK phosphorylation at Tyr 397; however 100 mM FAKi significantly reduced FAK phosphorylation among tested concentrations of 1, 10, and 100 mM. (D) Analysis of the integrated densities of the bands with the phosphor- to total FAK ratio. C represents untreated control and Cþ represents 100 nM fMLF treated only.

Article Snippet: Published by Wiley Periodicals, Inc. 448 JOURNAL OF ORTHOPAEDIC RESEARCH MARCH 2014 After 2 days, the explants were randomly distributed and were treated with fresh culture medium containing 10 or 100mM focal adhesion kinase inhibitor (FAKi) (Santa Cruz Biotechnology, Dallas, TX) to block phosphorylation of FAK at the kinase domain (Try 397) or were treated with fresh culture medium containing 10mM Src family kinase inhibitor (SFKi) (Selleckchem, Houston, TX) to block phosphorylation of SFKs at kinase domain (Tyr 416) for 2h.

Techniques: Western Blot, Phospho-proteomics, Control

Journal: Oncotarget

Article Title: CDCP1 is a novel marker of the most aggressive human triple-negative breast cancers

doi: 10.18632/oncotarget.11935

Figure Lengend Snippet: ( A ) MDA-MB-231 and BT-549 cells were transfected with a pool of 4 oligonucleotides (100 nM) that bind and degrade CDCP1 mRNA (see Materials and Methods for sequences) or with the appropriate negative control siRNAs. Cells were harvested at 48 h post-transfection, and CDCP1 expression was verified by western blot. Monoclonal anti-actin was used as a loading control. The knockdown efficiency was ≥ 60% for both CDCP1 forms. ( B ) CDCP1 siRNA-treated and control-treated MDA-MB-231 and BT-549 cells were analyzed for activation of CDCP1, Src, and PKCδ. Monoclonal anti-actin and anti-vinculin were used as loading controls.

Article Snippet: In the biochemical analyses, we used rabbit polyclonal antibody against CDCP1 (Merck Millipore); rabbit polyclonal phospho-CDCP1 (Tyr734) (Cell Signaling); mouse monoclonal anti-Src, clone GD11 (Merck Millipore); rabbit polyclonal phospho-Src family (Tyr416) (Cell Signaling); rabbit polyclonal anti-PKCδ (Cell Signaling); rabbit polyclonal phospho-PKCδ (Tyr311) (Cell Signaling); mouse monoclonal anti-vinculin, clone hVIN-1 (Sigma); anti-rabbit or -mouse IgG (GE Healthcare) as the secondary antibody; and peroxidase-linked mouse monoclonal anti-actin (Sigma-Aldrich).

Techniques: Transfection, Negative Control, Expressing, Western Blot, Activation Assay