Fig. 3: Spatio-temporal evolution of the spectrin clusters.
A Live imaging in dual mode (EPI and TIRF microscopy) of MEFs transiently transfected with GFP-βII-spectrin (green) and RFP-actin (magenta). Relevant frames are shown (scale bar 20 μm), as well as dynamic zooms of high-intensity βII-spectrin clusters only observed in the TIRF plane (white dashed boxes). Live images are representative of at least 3 independent experiments. B Temporal analysis of GFP-βII-spectrin clusters evolution (P0.95 of fluorescent signal intensity distribution). The resulting mask is shown with the clusters outlined, while the color-coded temporal projection is shown for the same cell presented in A, to highlight the dynamic nature of these clusters over time. C Auto-correlation coefficients between subsequent frames related to GFP-βII-spectrin (green) and RFP-actin (magenta) are shown (n = 8 independent cells and 486 frame pairs analyzed, data are presented as mean ± SD, statistical analysis paired t-test, ****p-value < 0.0001). D Representative frames during time lapse analysis of MEFs transiently transfected with GFP-βII-spectrin (green) and RFP-actin (magenta). The vectors highlight the orientation of the signal in the local window for the two channels. Coherency heatmaps are shown. E The graph reports the distribution of orientations of GFP-βII-spectrin (green) and RFP-actin (magenta), normalized to the Actin dominant direction (n = 8 cells in independent time lapses).
