YAP/TAZ-regulated NUAK2 enhances FA maturation and polarization, resulting in reduced cell motility. ECFCs were Triton-extracted concurrent with fixation for immunofluorescence of structural FAs. (A) Representative images of vinculin and paxillin visualized with Alexa Fluor 594 and 488 secondary, respectively. (B) Spatial differences in FA morphology were detected by subdividing cells into peripheral and central (5 µm from the edge of the cell) regions. FA length, an indicator of maturation, was found at the center and periphery of individual cells. (C) Vinculin⁺ FA length in the whole cell (P < 0.003), central region (P > 0.1), and peripheral region (P < 0.006), ANOVA with Tukey’s post hoc test for the whole cell and center and Kruskal-Wallis with Dunn’s post hoc test for the periphery. (D) Representative FA polarization distance indicated by the white line between the nucleus (red dot) and FA (white dot) intensity centroid. (E) FA polarization distance. P < 0.02; ANOVA with Tukey’s post hoc test. (F) ECFC wound closure rate after NUAK2 codepletion; n = 16; P < 0.006; ANOVA with Tukey’s post hoc test. (G) Immunoprecipitated MYPT1 (top), immunoblotted against total phosphorylated Serine (bottom), representative blot of n = 4 shown. (H) Quantification of p-Ser MYPT1/MYPT1; n = 4; P < 0.04; ANOVA with Tukey’s post hoc test. n.s., not significant. Repeated significance indicator letters (e.g., a–a) signify P > 0.05, while groups with distinct indicators (a vs. b) signify P < 0.05.