fak inhibitor 14 fib 14  (Tocris)

Journal: bioRxiv

Article Title: SIRPA suppresses integrin-dependent virus endocytosis

doi: 10.64898/2026.04.17.719277

Figure Lengend Snippet: A) U2OS cells were infected with TCRV (MOI=1) for 5 or 15 minutes. Western blots were probed with phospho-FAK (Y397) antibody. U, uninfected. B) U2OS cells were treated with FIB-14 for 1 hour and western blots were probed phospho-FAK (Y397) antibody. C) U2OS were treated with FIB-14 and infected with the indicated viruses. RNA was isolated at 24 hpi. Shown is the average ± SD of 3 (TCRV, JUNV C1) or 2 (LCMV) independent experiments. Student’s T test was used to determine significance. **, P ≤ 0.01; ****, P ≤ 0.0001. D) U2OS cells were treated with FIB-14 and infected with MACV, or VSV G pseudoviruses. Luciferase assays were performed at 48 hpi. Shown is the average ± SD of 3 independent experiments. One-way ANOVA was used to determine significance. ****, P ≤ 0.0001. E) shSIRPA or shCTRL cells were treated with FIB-14 and infected with the indicated viruses. RNA was isolated at 24 hpi. Shown is the average ± SD of 3 independent experiments. One-way ANOVA was used to determine significance. ***, P ≤ 0.001; ****, P ≤ 0.0001. F) VIAs were carried out with U2OS cells treated with FIB-14. Internalized viral RNA was analyzed by RT-qPCR. Shown is the average ± SD of 3 independent experiments. Student’s T test was used to determine significance. *, P ≤ 0.05; **, P ≤ 0.01.

Article Snippet: BTT-3033 and FAK inhibitor 14 (FIB-14) were purchased from TOCRIS, LHF-535 and favipiravir (6-fluoro-3-hydroxy-2-pyrazinecarboxamide) from TargetMol Chemicals and PP1 from Cayman Chemical.

Techniques: Infection, Western Blot, Isolation, Luciferase, Quantitative RT-PCR

Journal: bioRxiv

Article Title: SIRPA suppresses integrin-dependent virus endocytosis

doi: 10.64898/2026.04.17.719277

Figure Lengend Snippet: This step is inhibited by integrin inhibitors such as BT-3033 and the FAK inhibitor FIB-14. When virus binds to cells, an unknown signal also triggers Fyn to phosphorylate tyrosine residues in SIRPA’s ITIM region. This step is inhibited by SRC family inhibitors like PP1. SIRPA phosphorylation recruits and activates SHP2, which in turn dephosphorylates signaling molecules in the integrin signaling pathway like FAK and NMII. The dephosphorylation of integrin pathway components negatively impacts cytoskeletal rearrangement and actin polymerization, thereby reducing viral internalization. Originally created in BioRender. Yan, M. (2024) BioRender.com/h75b068 and modified by Ross, S. (2026) https://BioRender.com/ ooyrvzl.

Article Snippet: BTT-3033 and FAK inhibitor 14 (FIB-14) were purchased from TOCRIS, LHF-535 and favipiravir (6-fluoro-3-hydroxy-2-pyrazinecarboxamide) from TargetMol Chemicals and PP1 from Cayman Chemical.

Techniques: Virus, Phospho-proteomics, De-Phosphorylation Assay, Modification

Journal: bioRxiv

Article Title: Cell-supracellular structural relations solve the French Flag Problem without graded molecular control

doi: 10.1101/2025.10.24.683925

Figure Lengend Snippet: (A) Left, Sox9 levels increase over time until most limb progenitor cells adopt cartilage fate by 24 hours of culture. Right, Quantification of %Sox9+ area. (B) Left, DAPI and Sox9 in 24-hour ex vivo cultures plated at different densities show Sox9 activation is density dependent. Right, Quantification of %Sox9+ area. (C) Left, montage from live imaging of fibronectin in ex vivo cultures (Movie S3). Right, mean fibronectin intensity over time. (D) Focal adhesion components are enriched at interfaces between cells in 24-hour ex vivo cultures. (E) Ex vivo cultures treated with 10 µM FAKi display reduced Sox9 and FN and cell-cell associations. (F) Quantification of %Sox9+ area. *** p < .0001. (G) Sox9, Paxillin, FN, and Actin in 24-hour ex vivo cultures treated with FAKi, GSK205 (TRPV4 antagonist 50 µM), and GSK101 (TRPV4 agonist 15 µM). Sox9 images are from a separate replicate. (H) Quantification of %Sox9+ area. *** p = 0.0004; * p = 0.0110; ns, not significant. (I) Ex vivo limb progenitor cell culture stained for actin and Sox9. Scale bars, 100 µm (left) and 50 µm (right). (J) Sox9 and actin in ex vivo cultures treated with 10 µM Y27632. (K) Quantification of % Sox9+ area. **** p < .0001. (L) FN, Paxillin, Sox9, and Actin in ex vivo cultures treated with GSK205 (50 µM) and Y27632 (ROCK inhibitor, 10 µM). Right, quantification of % Sox9+ area. **** p < .0001. (M) Schematic showing cell-ECM structural formation guides cytoskeletal architecture toward the confirmation necessary for Sox9 activation. All violin plots show median and interquartile range; all scale bars = 50 µm; all error bars represent mean ± SD unless otherwise noted.

Article Snippet: The following pharmacological agents were utilized across cell culture assays: FAKi (FAK inhibitor 14, Tocris, Cat. 3414; 10 μM), Y-27632 (Tocris, Cat. 1254; 10 μM, TRPV4 inhibitor (GSK205, MedChemExpress, HY-120691A; 50 μM), TRPV4 activator (GSK101, Selleck, Cat. S8107; 15 μM), Wnt inhibitor (IWP-2, Tocris, Cat. 3533; 5 μM), MMP inhibitor (Batimastat, Tocris, Cat. 2961; 30 μM), Calmodulin inhibitor ( ) (W-7, Tocris, Cat. 0369; 1 μM).

Techniques: Ex Vivo, Activation Assay, Imaging, Cell Culture, Staining

Journal: bioRxiv

Article Title: Cell-supracellular structural relations solve the French Flag Problem without graded molecular control

doi: 10.1101/2025.10.24.683925

Figure Lengend Snippet: (A) DAPI, Actin, β1 integrin, and Vinculin in ex vivo cultures. Scale bar, 50 µm. (B) Sox9, Actin, and Paxillin in ex vivo cultures plated in a suspension of 3 mg/mL Matrigel and treated with 10 µM FAKi or 50 µM GSK205 to minimize cell loss in these treatment conditions. Scale bar, 50 µm. (C) DAPI, Sox9, and Actin in ex vivo cultures grown in Matrigel treated with 1 µM W7 (calmodulin inhibitor) show similar reductions as TRPV4 channel activity inhibition. Scale bar, 100 µm. (D) E5 limb bud section stained for DAPI, Sox9, and Actin shows a pattern of more diffuse actin fibers in Sox9-positive cells as compared to Sox9-negative cells. Scale bars, 150 µm (left) and 20 µm (right cropped images).

Article Snippet: The following pharmacological agents were utilized across cell culture assays: FAKi (FAK inhibitor 14, Tocris, Cat. 3414; 10 μM), Y-27632 (Tocris, Cat. 1254; 10 μM, TRPV4 inhibitor (GSK205, MedChemExpress, HY-120691A; 50 μM), TRPV4 activator (GSK101, Selleck, Cat. S8107; 15 μM), Wnt inhibitor (IWP-2, Tocris, Cat. 3533; 5 μM), MMP inhibitor (Batimastat, Tocris, Cat. 2961; 30 μM), Calmodulin inhibitor ( ) (W-7, Tocris, Cat. 0369; 1 μM).

Techniques: Ex Vivo, Suspension, Activity Assay, Inhibition, Staining

Journal: bioRxiv

Article Title: Hyperglycemic state and fluid shear stress affect metastatic breast cancer cell migration via focal adhesion kinase

doi: 10.1101/2025.05.22.655615

Figure Lengend Snippet: Co-centered cell migration tracks (a) and corresponding vector plots (b) for fluid flowed cells under high glucose without or with FAK inhibitor (FAK14). Fluid shear-induced MDA-MB-231 cell migration length under high glucose was significantly suppressed by FAK14 (c) (FF15: 15 dyne/cm 2 ). The migration inhibitory effects of FAK14 were also seen in confinement ratio (d) and arrest coefficient (e). Static MCF-10A cell data under low glucose were shown as controls for comparison. *, ***: p < 0.05 and 0.001, respectively. Bar graphs are plotted as mean ± SEM.

Article Snippet: For FAK inhibition studies, slides were placed in media containing 10 μM FAK inhibitor 14 (FAK14, Santa Cruz, sc-203950, Dallas, TX, USA) for 1 h. Then, slides were rinsed in phosphate buffered saline (PBS) before being placed in the flow chamber.

Techniques: Migration, Plasmid Preparation, Shear, Comparison

Journal: bioRxiv

Article Title: Hyperglycemic state and fluid shear stress affect metastatic breast cancer cell migration via focal adhesion kinase

doi: 10.1101/2025.05.22.655615

Figure Lengend Snippet: FAK inhibition by FAK14 significantly decreased MDA-MB-231 cell migration speed under fluid shear (FF15: 15 dyne/cm 2 ) and high glucose (a); t is the time in min after the fluid shear onset. Overall migration of the group was also blocked by FAK14 as in the RMS vs. t 1/2 plot (b). Motility coefficient was Static: 0.84, FF15: 1.76, and FF15-FAK14: 0.86, all under high glucose. ***: p < 0.001, Bar graphs are plotted as mean ± SEM.

Article Snippet: For FAK inhibition studies, slides were placed in media containing 10 μM FAK inhibitor 14 (FAK14, Santa Cruz, sc-203950, Dallas, TX, USA) for 1 h. Then, slides were rinsed in phosphate buffered saline (PBS) before being placed in the flow chamber.

Techniques: Inhibition, Migration, Shear

Journal: bioRxiv

Article Title: Hyperglycemic state and fluid shear stress affect metastatic breast cancer cell migration via focal adhesion kinase

doi: 10.1101/2025.05.22.655615

Figure Lengend Snippet: (a) Initial scratch area at 0 h and the area imaged at 24 h under low (L) and high (H) glucose without or with FAK inhibitor (FAK14). There was no appreciable difference in scratch wound area with low (L) and high (H) glucose conditions. On the other hand, FAK14 considerably suppressed the covering of the scratch (b) and movement of cells towards the scratch (c). **: p < 0.01 (n = 5). Bar graphs are plotted as mean ± SEM.

Article Snippet: For FAK inhibition studies, slides were placed in media containing 10 μM FAK inhibitor 14 (FAK14, Santa Cruz, sc-203950, Dallas, TX, USA) for 1 h. Then, slides were rinsed in phosphate buffered saline (PBS) before being placed in the flow chamber.

Techniques:

Journal: bioRxiv

Article Title: Hyperglycemic state and fluid shear stress affect metastatic breast cancer cell migration via focal adhesion kinase

doi: 10.1101/2025.05.22.655615

Figure Lengend Snippet: (a) Images of transwell migration assay for MDA-MB-231 cells under low (L) and high (H) glucose without or with FAK inhibitor (FAK14). Arrows example the regions of stained cells. (b) The invasive cell count was not affected by glucose concentration, while FAK14 substantially decreased cell movement across the membrane. Invasive cell count is presented as % of low glucose control without FAK14. *: p < 0.05 (n = 5). Bar graphs are plotted as mean ± SEM.

Article Snippet: For FAK inhibition studies, slides were placed in media containing 10 μM FAK inhibitor 14 (FAK14, Santa Cruz, sc-203950, Dallas, TX, USA) for 1 h. Then, slides were rinsed in phosphate buffered saline (PBS) before being placed in the flow chamber.

Techniques: Transwell Migration Assay, Staining, Cell Counting, Concentration Assay, Membrane, Control