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. 2021 Mar 18;220(4):e202003052. doi: 10.1083/jcb.202003052

Figure 6.

Figure 6.

The FLS tip complex actin regulatory protein is semidynamic. (A) Time course recovery of AF488 actin (n = 65), GFP-Fascin (n = 40), AF488 TOCA-1(n = 51), GFP-Diaph3 (n = 34), AF488 N-WASP (n = 39), pmKate-GBD (n = 39), AF488 Ena (n = 36), and AF568 VASP (n = 51) after photobleaching in tips of FLSs at steady state grown for at least 30 min. Solid lines show the median, and the shaded area is the 95% confidence interval. (B) The half-time and percentage recovery from fitted exponential curves for each protein. Error bars represent the 95% confidence interval of the mean. (C) Actin regulatory protein localization to FLS and fluorescence recovery after photobleaching at FLS tips (green: protein of interest, magenta: AF647 actin). Protein concentrations unlabeled/labeled: Actin 210/4,000; TOCA-1 10/4; VASP 45/24; N-WASP 18/18; GBD 2.46/0; fascin 650/625; Diaph3 16/15; and Ena 60/60 in nanomolars. Scale bars = 2 µm (main images) or 1 µm (insets). (D) Time series cross-correlation of the absolute values of the FLS growth velocity and background-corrected N-WASP (n = 780), Ena (n = 2,003), and VASP (n = 2,783) intensities at FLS tip complexes, averaged over all measured FLS trajectories. The graph shows the cross-correlation coefficient at the given time shift (negative time shifts mean that intensity changes precede velocity changes). Shaded areas are 95% confidence interval of the mean.