Journal: bioRxiv

Article Title: Focal adhesions are controlled by microtubules through local contractility regulation

doi: 10.1101/2023.04.17.535593

Figure Lengend Snippet: (A) Representative images of Vinculin-mIFP-transfected HT1080 cells carrying the OptoKANK constructs before and after blue light illumination of the focal adhesion (yellow dotted line) for control cells, cells treated with a calpain inhibitor, calpastatin, a kinesin-V inhibitor, monastrol, a dynamic inhibitor, dynasore, a MMP inhibitor, Batimastat, a MAP4K4 inhibitor, MAP4K4-IN-3, and depleted for BNIP2 and ARP2. Graph shows the normalized mean vinculin intensity after the illumination of cells treated as indicated (Data are presented as mean ± s.e.m; Ctrl, n =19; Nocodazole, n = 12; Calpastatin, n = 10; Monastrol, n = 8; Dynasore, n = 10; MMP inhibitor, n = 7; CNO3, n = 8; Tubacin, n = 12, MAP4-K4-IN3, n = 9; siRNA BNIP2, n = 16; siRNA AR2, n = 10). Immunoblots of BNIP2, ARP2, acetylated tubulin and GAPDH are shown in the black box.

Article Snippet: Pharmacological treatments were performed using the following concentrations of inhibitors or activators: 1 μM for Nocodazole (Sigma-Aldrich), 0.4 μM for Y-27632 dihydrochloride (Sigma-Aldrich), 20 mM for Acetyl-Calpastatin (Tocris, Cat. No. 2950), Monastrol (Merk, cat. No. 254753-54-3), 80 µM for Dynasore (Abcam Cat. No. 120192), Batimastat (Tocris Cat. No. 2961), Tubacin (Cat. No. 3402), MAP4K4-IN-3 (MedChemExpress, Cat. No.: HY-125012), 1 µM for FRAX1036 (MedChemExpress Cat. No. HY-19538), 50 µM for Kinesore (Cat. No. 6664), 5 µM for PF-57328 (Tocris, Cat. No. 3239), and 1 μg ml - for Rho activator II (CNO3, Cytoskeleton).

Techniques: Transfection, Construct, Western Blot