fak inhibitor 14 faki  (Tocris)

Journal: Hormones & Cancer

Article Title: Thyroid Hormone Controls Breast Cancer Cell Movement via Integrin αV/β3/SRC/FAK/PI3-Kinases

doi: 10.1007/s12672-016-0280-3

Figure Lengend Snippet: T3 regulates Src/FAK/PI3K complex formation. T-47D breast cancer cells were exposed for 20 min to T3 (1 nM) in the presence or absence of the Src inhibitor (PP2 10 μM), the PI3K inhibitor wortmannin (WM 30 nM), FAK inhibitor (FAKi 1 μM), or the MAPK inhibitor PD98059 (PD 5 mM). a, b Tyr397- FAK phosphorylation was evaluated through Western blot analysis. c Breast cancer cells treated with T3 for 20 min in the presence or absence of FAK inhibitor and the inhibitor of the extracellular regulated kinase 1/2 (ERK1/2), PD98059. Cell contents of total or phosphorylated ERK1/2 are shown. d, e T-47D breast cancer cells were exposed to T3 (1 nM) for 20 min in the presence or absence of PP2, WM, and FAKi. Cell protein extracts were immunoprecipitated with an antibody vs. FAK (d) and p85alpha (e). IPs were assayed for co-immunoprecipitation with Src, FAK, and p85α. Membranes were re-blotted for the immunoprecipitated protein to show equal input. The experiments were performed in triplicate; representative images are shown. CON control

Article Snippet: Before experiments investigating non-transcriptional effects, cancer cells were kept in medium containing no FBS for 8 h. T3, PD98059 (PD), wortmannin (WM), and LY294002 (LY) were from Sigma-Aldrich (Saint Louis, MO); 4-amino-5-(4-chlorophenyl)-7-(t-butyl) pyrazolo (3,4-d) pyrimidine (PP2) was from Calbiochem (La Jolla, CA) and Tetraidothyroacetic acid (Tetrac) and FAK inhibitor (FAKi) from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Phospho-proteomics, Western Blot, Immunoprecipitation, Control

Journal: Hormones & Cancer

Article Title: Thyroid Hormone Controls Breast Cancer Cell Movement via Integrin αV/β3/SRC/FAK/PI3-Kinases

doi: 10.1007/s12672-016-0280-3

Figure Lengend Snippet: T3 modulates breast cancer cell adhesion and migration through Src/FAK/PI3K pathway. Cells were treated with T3 (1 nM) for 48 h in the presence or absence of the Src inhibitor (PP2; 0.2 μM), the PI3kinase inhibitor wortmannin (WM; 30 nM), the FAK inhibitor (FAKi 1 μM), and MAP Kinase inhibitor PD98059 (PD 5 mM). a, b The arrows indicate the direction of migration. The upper black lines indicate the starting line and the lower black lines the mean migration distance. c, d Representative images of T-47D cell adhesion to gelatin after T3 treatment (1 nM/2 h) are shown. Experiments were performed in triplicate; representative images are shown

Article Snippet: Before experiments investigating non-transcriptional effects, cancer cells were kept in medium containing no FBS for 8 h. T3, PD98059 (PD), wortmannin (WM), and LY294002 (LY) were from Sigma-Aldrich (Saint Louis, MO); 4-amino-5-(4-chlorophenyl)-7-(t-butyl) pyrazolo (3,4-d) pyrimidine (PP2) was from Calbiochem (La Jolla, CA) and Tetraidothyroacetic acid (Tetrac) and FAK inhibitor (FAKi) from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Migration