CAFs form larger adhesions that turn over more slowly than NFs do. (A) IF staining of vinculin (green) and actin (red) in NFs and CAFs. White boxes indicate selected areas of interest in the zoom boxes. Bars: (original images) 25 µm; (zoomed images) 5 µm. (B and C) Box plots show mean adhesion size (μm2; B) and adhesion number (C) in NFs and CAFs. More than 600 adhesions (B) and than 50 cells (C) per condition were analyzed from three independent experiments. ***, P < 0.001, determined by Mann Whitney U test. (D) Time-lapse images show adhesion assembly and disassembly. NFs and CAFs were transfected with vinculin–GFP (pseudo-colored in gold; white arrowheads, adhesion assembly; blue arrows, adhesion disassembly; Videos 9 and 10). Bar, 10 µm. (E and F) Quantification of the apparent t1/2 of adhesion assembly (E) and disassembly (F) for NFs and CAFs. 50–60 adhesions were analyzed per condition from four independent experiments. ***, P < 0.001, determined by Mann Whitney U test. All box plots range from the 25th to the 75th percentiles; the central line indicates the median, and the whiskers range from the 5th to the 95th percentiles.
