Fig. 7: DAFS variant VcnTSs exhibit loss of Vcn force-stabilized dynamics.
From: Molecular basis and cellular functions of vinculin-actin directional catch bonding
a, b Representative Vcn−/− VcnTS and Vcn−/− VcnTS E1015A-E1021A FAs undergoing FRAP time lapse. Scale bar: 2 μm c FRAP recovery curves are shown for cells expressing VcnTS and VcnTS E1015A-E1021A (n = 25, 22 from N = 11 independent experimental days). Measure of center for error bars is average of n normalized intensities at respective timepoint for VcnTS or VcnTS E1015A-E1021A expressing cells. Exponential fit curves are fitted to averages at respective timepoints. d Box-whisker plots of mobile fraction of FAs analyzed in c, using two-tailed Welch’s t-test for unequal variances (p = 0.0000054) e Box whisker plots of FRAP recovery half-time of FAs analyzed in c, using one-way ANOVA followed by F-test (p = 0.25). Box depicts the median as the center value, the 25th percentile as the lower bound, and the 75th percentile as the upper bound. Whiskers extend 1.5 times the IQR from the bottom and top of box, or to the minimum and maximum of the data if the data does not extend to the whiskers. Values outside the whiskers are plotted as individual points. f Correlation between FRAP recovery half-time and FRET efficiency for VcnTS corresponds to the force-stabilized state. g Correlation between recovery half-time and FRET efficiency for VcnTS E1015A-E1021A corresponds to the force-independent state. h Regression slopes calculated for panel (f–g). A least-squares linear regression was fit to the FRAP half-time and FRET efficiency data for each construct. Bars represent linear regression slope, and error bar represents standard error of the regression slope. Star indicates slope is statistically significant from zero as detected by one-way ANOVA followed by an F-test (p = 0.007 for VcnTS; p = 0.49 for VcnTS E1015A-E1021A). Source data are provided as a Source Data file.
