α-Catenin regulates adhesion maturation. (A) Immunoblot for α-catenin showing KD in NIH3T3 cells. (B) Immunostaining for vinculin and α-catenin in WT and α-catenin KD NIH3T3 cells. (C) Quantification of cell area and number of mature adhesions per square micrometer in WT and α-catenin KD NIH3T3 cells. n = 17 cells in each case. (D) Immunostaining for zyxin shows the presence of mature adhesions in WT MEFs and α-catenin KD cells expressing WT GFP-α-catenin (white arrows), but only the presence of focal complexes in α-catenin KD cells and α-catenin KD cells expressing GFP-α-catenin L344P (yellow arrows). (E) Formation of sliding adhesions (arrows) on Matrigel-coated coverslips by WT cells, α-catenin KD cells, and α-catenin KD cells expressing WT GFP-α-catenin or GFP-α-catenin L344P (n > 15 cells in each case). (F) Immunoblot for α-catenin in WT MEFs and α-catenin KD cells expressing GFP-α-catenin L344P, GFP-α-catenin L344A, and GFP-α-catenin V796A. Statistical analysis for the NIH3T3 cell area and number of adhesions was performed with two-tailed unpaired t test followed by Welch’s correction. Statistical analysis of the number of sliding adhesions was performed by ANOVA followed by Tukey’s post hoc test correction (*, P < 0.05; **, P < 0.01). Source data are available for this figure: SourceData FS4.