primary antibodies against p fak  (Cell Signaling Technology Inc)

Journal: Cancers

Article Title: Anticancer Effect of Heparin–Taurocholate Conjugate on Orthotopically Induced Exocrine and Endocrine Pancreatic Cancer

doi: 10.3390/cancers13225775

Figure Lengend Snippet: Inhibition effect of LHT on the cellular proliferation and migration through VEGF receptor, ERK and FAK pathways. ( A ) Multiple signaling pathways from the VEGF-A/VEGFR2 interaction to induce cellular migration and proliferation. LHT inhibits VEGF-A/VEGFR2 binding through VEGF-A/LHT interactions. ( B ) Western blot (cropped blots) of cell lysate of HUVECs after cultivation with VEGF (10 ng/mL) or VEGF with LHT (100 µg/mL) for 24 h. Full-length blots were presented in . ( C ) Quantitative analysis of Western blotting bands cell lysate of HUVECs after cultivation with VEGF (10 ng/mL) or VEGF with LHT (100 µg/mL) for 24 h. Data were expressed with mean ± s.e.m. (n = 3). * p < 0.05, *** p < 0.001 versus the VEGF-treated group.

Article Snippet: Then, Western blotting was carried out for checking the phosphorylation of VEGFR, ERK1/2 and FAK molecules with primary antibodies against ERK1/2 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), p-ERK1/2 (Santa Cruz Biotechnology), FAK (Signalway Antibody LLC., College Park, MD, USA), p-FAK (Signalway Antibody LLC), VEGFR (Abcam), p-VEGFR (Abcam) followed by incubation with horseradish peroxidase-conjugated secondary antibodies.

Techniques: Inhibition, Migration, Protein-Protein interactions, Binding Assay, Western Blot

Journal: The Journal of Cell Biology

Article Title: Src- and confinement-dependent FAK activation causes E-cadherin relaxation and β-catenin activity

doi: 10.1083/jcb.201706013

Figure Lengend Snippet: FAK activity is required and activated by Src under stimulation. (A) FAK kinase activity FRET biosensor. The higher the activity, the lower the FRET index. (B) FRET index (averaged over the whole cell) of the stably expressed FAK biosensor in unstimulated ( n = 593), 4-h HGF-treated ( n = 183), PF228-treated ( n = 455), PP1-treated ( n = 392) cells, single cells ( n = 74), and 4-h HGF-treated single cells ( n = 108), confluent cells ( n = 91), and 4- to 5-h HGF-treated confluent cells ( n = 202). ns, not significant. (C) Top: FAK domain organization and tyrosine sites. Bottom: Typical Western blot from lysates of unstimulated and HGF-treated (3 h) with or without PP1 or PF228. (D) Phosphorylation levels FAK Y397, Y576/577, Y861 normalized to total FAK level from Western blots of cells treated as in panel c ( n = 7 lysates). (E) Top: Typical micrographs of GFP-β-catenin cells with and without HGF and PF228. Bottom: Area after 2 h normalized by the area at t = 0 for unstimulated cells, HGF-treated (data from Fig. S1 I), and HGF- and PF228-treated cells ( n = 10 cells). (F) GFP intensity in the nucleus (relative to cytoplasm) of GFP-β-catenin 5 h with or without HGF (50 ng/ml; data from ), and with PF228 (10 µM) with or without HGF ( n = +/− 37/43 cells). (G) FRET index change of EcadTSMod in cells 5 h with and without HGF (50 ng/ml; data from ), and with PF228 (25 µM) with or without HGF ( n = +/− 110/99 cells). (H) Top: Typical micrographs of wounded sheet migration of GFP-β-catenin cells with and without (same control as in ) PF228 (25 µM). Bottom: Migration speed (mean distance traveled by cell front per hour) with or without (same control as ) PF228 (25 µM; n = 121 cells). Bars: (wound healing) 100 µm; (FAK sensor) 20 µm. Two-tailed Mann–Whitney or Kruskal–Wallis tests. Values plotted are mean ± SEM.

Article Snippet: Proteins were transferred to nitrocellulose (Whatman), blocked, and probed with primary antibodies against FAK, p-FAK Y397, p-FAK Y576/577, p-FAK Y86 (610088; BD Sciences; 11765; Santa Cruz Biotechnologies; 3281P; Cell Signaling Technology; and 44626; Invitrogen, respectively) or primary antibodies against E-cadherin, β-catenin, or α-tubulin (clone 36-610181; BD Sciences; CM1181; ECM Biosciences; and T9026; Sigma-Aldrich).

Techniques: Activity Assay, Stable Transfection, Western Blot, Phospho-proteomics, Migration, Control, Two Tailed Test, MANN-WHITNEY