Figure 6.

FP mechanical switching in VinTS is sensitive to manipulation of the vinculin-actin catch bond

(A–Y) Representative vinculin^−/− MEFs expressing VinTS-I997A (A–E), VinTS (F–J), VinTS-E1015A (K–O), VinTS- E1021A (P–T), or VinTS-E1015A-E1021A (U–Y) on FN-coated glass, showing images of acceptor intensity, donor intensity, FRET efficiency in the FA mask, stoichiometry in the FA mask, and an ES-histogram of FA-masked pixels for the cell. For single-cell ES-histograms in (E), (J), (O), (T), and (Y), the color bars indicate pixel counts.

(Z) ES-histograms for whole-cell populations (N = 59/89/92/88/101 cells over 2/4/3/3/3 experimental days for VinTS-I997A/VinTS/VinTS-E1015A/VinTS-E1021A/VinTS-E1015A-E1021A), where color bars indicate the cell-averaged fraction of FA-masked pixels in each bin.

(AA) Boxplot of fraction of pixels in each cell in the low $E$, high $S$ bin ($E<0.15,S>0.60$) for each construct. Differences between groups were detected using the Steel-Dwass test. Levels not connected by the same letter are significantly different at p < 0.05. p values are given in Note S3.

The data are a new analysis of the 3-channel FRET images from an experiment in a previous publication.³⁹

See also Figures S19–S21.