Journal: Materials Today Bio

Article Title: Modulating cell surface chemistry through mild reduction reinforces extracellular-to-intracellular transmission forces and mechano-signaling

doi: 10.1016/j.mtbio.2026.102908

Figure Lengend Snippet: TCEP-induced mild reduction of the cell surface promotes early adhesion–associated FAK activation and focal adhesion assembly. (A) Schematic illustration of the experimental workflow for monitoring FAK activation using a Lyn–FAK FRET biosensor. This schematic was created using BioRender.com . (B) Representative time-lapse FRET ratio images showing ECFP/YPet FRET ratios during early adhesion (Scale bar = 20 μm). (C) Quantification of the time-dependent ECFP/YPet FRET ratio during early adhesion. Data are presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 compared with the Ctrl group. (D) Representative immunofluorescence staining for F-actin (red) and pFAK (green) (Scale bar = 100 μm). (E-J) Quantitative image analysis of cell morphology and focal adhesion (FA) parameters (Scale bar = 50 μm). Data are presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: The Lyn–FAK FRET biosensor plasmid (Plasmid #78299, Addgene, Watertown, MA, USA) was inserted into the pAd/CMV/V5-DESTTM GatewayTM vector (V49320, Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer's instructions to generate an adenoviral expression construct.

Techniques: Activation Assay, Immunofluorescence, Staining

Journal: Science Advances

Article Title: Mechanochemical waves in focal adhesions during cell migration

doi: 10.1126/sciadv.adw6425

Figure Lengend Snippet: ( A ) The FAK biosensor is composed of ECFP, SH2 domain, flexible linker, FAK substrate peptide, YPet, and FAT domain. ( B ) Schematics illustrating the FRET effect of the FAK biosensor upon the actions of FAK phosphorylation or dephosphorylation. Upon phosphorylation of Y397 in the biosensor FAK substrate peptide, the SH2 domain forms an intramolecular complex with the phosphotyrosine side chain, increasing the distance between the FRET pair to alter the FRET signal. Dephosphorylation reverses the FRET change. ATP, adenosine 5′-triphosphate. ( C ) C-terminal FAT domain recruits the biosensor to FAs. YPet intensity showing slight changes before and after FAK inhibition (FAKi; 10 μM PF-573228, 60 min). ( D ) ECFP/FRET signal before and after FAKi (10 μM PF-573228, 60 min) showing that the biosensor is specific and sensitive to FAK activity. ( E ) EFCP/FRET signal at individual FAs [ n = 195 FAs from seven cells across three independent experiments for FAKi (10 μM PF-573228, >60 min); n = 151 FAs from six cells across three independent experiments for DMSO control; means ± SD]. ( F ) Fluorescence lifetime image and quantification for fibroblasts expressing the FAK biosensor ( n = 23 FAs from six cells across three independent experiments). Scale bar, 20 μm. ( G ) FRET efficiency image and quantification for FAs and cytosol ( n = 19 FAs from five cells across three independent experiments). Scale bar, 20 μm. a.u., arbitrary units.

Article Snippet: The FAK FRET biosensor construct is available from Addgene (plasmid no. 78303).

Techniques: Phospho-proteomics, De-Phosphorylation Assay, Inhibition, Activity Assay, Control, Fluorescence, Expressing