Motility assay was performed by using a chamber containing monolayer-coated polyethylene teraphthalate membrane (six-well insert, pore size of 8 μm; Becton Dickinson, Franklin Lakes, NJ, USA). Both scramble and MUC5AC knockdown cells of A549 (2 × 106 cells per well) and H1437 (1.5 × 106 cells per well) were seeded in six-well plates. After 24 h incubation period, the migrated cells that had reached the lower chamber were stained with a Diff-Quick stain set and counted in different fields. The average number of migrated cells per representative field was calculated. Similarly, for FAK inhibitor treatment studies, cells were seeded along with FAK inhibitor (FAK inhibitor 14, Cayman chemical, CAS4506-66-5), which direct inhibitor and specific for FAK (Y397)36 (link) and incubated for 24 h and migrated cells were calculated.
Fak inhibitor 14
Manufactured by Cayman Chemical
Sourced in United States
FAK inhibitor 14 is a small molecule compound that inhibits the activity of Focal Adhesion Kinase (FAK), a protein that plays a critical role in cell signaling pathways. This compound can be used as a research tool to study the biological functions of FAK and its involvement in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Ly294002, by Cayman Chemical (1 mentions)
Hbmec, by Thermo Fisher Scientific (1 mentions)
Neon transfection system, by Thermo Fisher Scientific (1 mentions)
Pspax2, by Addgene (1 mentions)
Pmd2 g, by Addgene (1 mentions)
Endothelial cell medium (ecm), by ScienCell (1 mentions)
Mg132, by Cayman Chemical (1 mentions)
3 protocols using fak inhibitor 14
1
Motility Assay for MUC5AC Knockdown
2
Inhibition of FAK and PI3K Signaling
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FAK inhibitor (FAK inhibitor 14; Cayman Chemical, Ann Arbor, MI, USA) and PI3K inhibitor (LY294002; Cayman Chemical) were dissolved in dimethyl sulfoxide (Sigma) and used at appropriate concentrations.
3
Lentiviral Transduction and Autophagy Induction in Human Brain Microvascular Endothelial Cells
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Human brain microvascular endothelial cells (HBMECs), obtained from Cell Systems (ACBRI 376, Kirkland, WA, USA), were cultured in endothelial cell medium (#1001, ScienCell Research Laboratories, Carlsbad, CA, USA). Inducible gene expression in HBMECs was achieved through lentiviral infection. Lentiviruses were generated in HEK293T cells with the assistance of two other plasmids, pMD2.G (#12259, Addgene) and psPAX2 (#12260, Addgene). For siRNA-mediated knockdown experiments, siRNAs were introduced into HBMECs using standard electroporation techniques (Neon Transfection System, Invitrogen, Carlsbad, CA, USA). Autophagy was induced by treating HBMECs with Earle’s Balanced Salt Solution (EBSS) for 2 h, followed by LC3 puncta or p62 turnover assays. All chemicals, including pepstatin/E-64D, MG132, and FAK inhibitor 14, were purchased from Cayman Chemical.
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