anti-pfak tyr925 (cat# 3284) antibody  (Cell Signaling Technology Inc)

Journal: Frontiers in Genetics

Article Title: Exploring the prognostic significance of lactate-mitochondria-related genes in prostate cancer

doi: 10.3389/fgene.2024.1515045

Figure Lengend Snippet: MPO inhibits prostate cancer metastasis. (A) The cell number of LNCaP cells was counted at different time points. (B, C) Cell cycle analysis through PI staining was detected by FACS (B) , and the cell cycle distribution was presented (C) . (D, E) LNCaP cells were analyzed for cell migration and invasion. Representative images of crystal violet-stained migrated (D) or invaded (E) cells are presented (scale bar, 100 μL). (F) The Tyr925-phosphorylated level of FAK and total FAK, E-Cadherin, and Snail in LNCaP cells were detected by Western blot analysis. Actin was used as a loading control. All data are presented as the mean ± SD of three independent experiments; ns, not significant.

Article Snippet: Anti- FAK (Cat# 3285), anti-pFAK Tyr925 (Cat# 3284), anti-Snail (Cat# 3879), anti-E-Cadherin (Cat# 3195) and anti-actin (Cat# 4970) antibodies were purchased from Cell Signaling Technology.

Techniques: Cell Cycle Assay, Staining, Migration, Western Blot, Control

Journal: Cancer Research Communications

Article Title: Clotting Promotes Glioma Growth and Infiltration Through Activation of Focal Adhesion Kinase

doi: 10.1158/2767-9764.CRC-24-0164

Figure Lengend Snippet: Growth of GBM cells in fibrin clot and plasma clot depends on integrins β1 and β3. A, Integrins β1, β3, β5, and αV as well as total and activated FAK at Y397 and Y925 were analyzed in subconfluent extracts from U87MG, U373MG, and U343MG GBM cells by immunoblotting. α-tubulin served as a loading control. B, Western blot analysis of integrin β1 and integrin β3 expression and total FAK and FAK activation at Y925 in extracts from U87MG and U373MG cells 3 days after transfection with siRNA against integrins β1 (siβ1) and β3 (siβ3) compared with treatment with control siRNA (siCtrl; B , top). FAK expression after transfection with siRNA against FAK (siFAK) compared with siCtrl ( B , bottom). C–F, Fold increase in cell proliferation was analyzed over time in fibrin clot–embedded ( C ) and plasma clot–embedded ( D ) U87MG cells and in fibrin clot–embedded ( E ) and plasma clot–embedded ( F ) U373MG cells following transfection with siβ1, siβ3, and siRNA against FAK compared with siCtrl by phase-contrast microscopy. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 compared with siCtrl.

Article Snippet: The membrane was blocked with 5% non-fat dry milk or 5% BSA in PBS containing 0.1% Tween 20 for 1 hour at room temperature and probed for anti-integrin β1 (Cell Signaling Technology, catalog # 9699, RRID: AB_11178800), anti-integrin β3 (BD Biosciences, catalog # 611140, RRID: AB_398451), anti-integrin β5 (Cell Signaling Technology, catalog # 3629, RRID: AB_2249358), anti-integrin αV (BD Biosciences, catalog # 611012, RRID: AB_398325), anti-FAK (Cell Signaling Technology, catalog # 3285, RRID: AB_2269034), anti-pFAK Y397 (Thermo Fisher Scientific, catalog # 700255, RRID: AB_2532307), anti-pFAK Y925 (Cell Signaling Technology, catalog # 3284, RRID: AB_10831810), or α-tubulin (Sigma-Aldrich, catalog # T-6199, RRID: AB_477583) overnight at 4°C.

Techniques: Clinical Proteomics, Western Blot, Control, Expressing, Activation Assay, Transfection, Microscopy