Extended Data Fig. 7: Fast directed motions of FMNL2 inside and outside the lamellipodium.
a, Fluorescence image of α-actinin-GFP in a MEF co-transfected with FMNL2-mEos2 (left). Kymographs generated from a raw sptPALM acquisition (50 Hz) to measure FMNL2-mEos2 fast directed motions occurring in the lamellipodium (1) or outside the lamellipodium (2,3) (left panel, dashed lines). Fast motions of FMNL2-mEos2 highlighted (white lines) and corresponding velocities (nm.s-1). b, Distributions of speed for fast directed motions occurring in the lamellipodium (n = 29 events), or outside the lamellipodium (n = 38 events) for FMNL2-mEos2 (pooled over 3 cells from 2 experiments), box plots displaying median (notch) and mean (square) ± percentiles (25-75%). c, Super-resolution intensity image of FMNL2-mEos2 in the a MEF lamellipodium obtained by sptPALM (50 Hz, duration: 150 s) (left). Right, FMNL2-mEos2 kymographs generated from a sptPALM time-lapse sequence (2 Hz) along LOI corresponding to directed motions occurring within or outside the lamellipodium. d, same as c for FMNL2-I704A-mEos2. e, same as c for FMNL2-A272E-mEos2. f, density of directed motions, box plots displaying median (horizontal line) and mean (square) ± percentiles (25-75%) for each cell. Data pooled from independent experiments: mEos3.2-Sra1, n = 5 cells (pooled over 2 experiments); mEos3.2-Sra1-WCA*, n = 4 cells (over 2 experiments) and mEos3.2-Sra1-AD, n = 6 cells (over 3 experiments). Images in a, c-e are representative of 2, 2, 2 and 3 independent experiments that yielded similar results. Where indicated, statistical significance were obtained using two-tailed Mann-Whitney rank sum test for speed (b) and density (f) of directed motions. Also see Supplementary Table 3. Numerical source data are provided in Source data.
