Fig. 2: Perinuclear actin and nuclear abnormalities are present in MET hyperactivated cells and completely reverted after MET ablation.

a Actin cytoskeleton and nucleus visualization in LoVo MET + /MET-KO fixed cells through Phalloidin (red) and DAPI (blue) staining. White arrows indicate the actin patches observed in MET+ cells. Scale bar: 20 μm. b Confocal 3D rendering of cells fixed and stained as in (a). White arrows indicate the actin patches observed in MET+ cells. c Nucleus and actin visualization in LoVo MET + /MET-KO cells fixed after 12/18/24/48 h from seeding. White arrows points at the actin patches. A quantification of the number of actin patches observed per field and normalized on DAPI is provided for each time point. 10 fields per condition were analyzed (181 and 135 cells for MET+ and MET-KO samples respectively). Data are reported as mean ± SEM. Statistic was calculated by Two-way ANOVA. Scale bar: 20 μm d Confocal 3D rendering of representative nuclei from experiment in (c). Nuclear height quantification was performed on 10 fields for each time point as in (c) by analyzing Z-stacks images. Data are shown as mean ± SEM. Statistic was calculated by Two-way ANOVA. Scale bar: 20 μm. The scale bar refers to the xy-plane. e Transmission electron microscopy of LoVo MET+ cells. White arrows point at actin filaments. Scale bars: left 5 μm, right 1 μm. f ODT and 3D reconstruction of LoVo MET+ aberrant actin patches. Z-depth coding is reported as a scale of colors. g ODT time lapse of LoVo MET+ cells showing actin patches generation. Pictures of cells were acquired every 10’ for 12 h. Full video is provided as Supplementary video 3. Scale bar: 10 μm.