focal adhesion kinase inhibitor fak inhibitor 14  (Millipore)

Journal: Journal of Biological Chemistry

Article Title: Tumor Necrosis Factor-α (TNFα)-induced Ceramide Generation via Ceramide Synthases Regulates Loss of Focal Adhesion Kinase (FAK) and Programmed Cell Death

doi: 10.1074/jbc.m115.658658

Figure Lengend Snippet: FIGURE 8. The inhibition of the autophosphorylation of FAK on Tyr-397 by FAK inhibitor 14 (Y15) is sufficient to trigger cell death. MCF-7 cells were treated with a 10 M concentration of the focal adhesion kinase inhibitor Y15 for the time indicated. A, the levels of phosphorylated and total FAK and pro-caspase-7 were analyzed by Western blot upon Y15 treatment for the indicated time. A representative experiment is shown. Similar results were obtained in at least two additional experiments. GAPDH was used as a gel loading control. B, cell viability was measured by the MTT assay. C, plasma membrane permeabilization was measured by the LDH release assay. D, a caspase-3/7-like DEVDase activity assay was performed in the time indicated. E, MCF-7 cells were pretreated with 10 M z-DEVD for 1 h and then treated with Y15 for 6 h. Plasma membrane permeabilization was measured by LDH release into the medium. Data represent mean and S.E. (error bars) of at least three independent experiments. B–E, statistical significance was determined by one-way ANOVA with Bonferroni post-test. ***, p 0.001; ****, p 0.0001 versus vehicle.

Article Snippet: Anti-PARP-1 and focal adhesion kinase inhibitor 14 (Y15) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Inhibition, Concentration Assay, Western Blot, Control, MTT Assay, Clinical Proteomics, Membrane, Lactate Dehydrogenase Assay, Activity Assay