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. 2022 Jul 19;10:959521. doi: 10.3389/fcell.2022.959521

FIGURE 2.

FIGURE 2

DAPK1 regulates adhesion growth and force production. MEFs treated with DAPK1 inhibitor or stable DAPK1 shRNA MEFs were fixed after spreading on fibronectin-coated glass dishes for 30 min, 60 min or 16 h, followed by anti-paxillin/AlexaFluor 488 immunostaining. (A) Micrographs showing the distribution of paxillin in cells fixed after spreading on fibronectin-coated glass for 30 min. Scale bar: 10 μm. (B) Quantification of the adhesion sizes (n > 400 adhesions from >10 cells in each case, mean ± SEM). ***p < 0.001. (C) Average area ± s.e.m of the cells (n > 10 cells in each case). ***p < 0.001. Experiment was repeated three times. (D) Displacements (red arrows) of pillars that show the contractile pair (green arrows marked with yellow oval) at the periphery of a spreading MFF during phase 2 of cell spreading (Dobereiner et al., 2004). Scale bar: 1 μm. (E) Displacements versus time of contractile pillars in MEFs treated with or without DAPK1 inhibitor. (F) Mean ± SEM of the T1/2 (time of contraction above half-maximal displacement) distributions of pillar displacements by MEFs during phase 2 of cell spreading before and after treatment with DAPK1 inhibitor. ***p < 0.001. N > 30 pillars in each case. Experiment was repeated twice.