Fig. 3: MET knock-out in MET dependent cells prompts proper organization of the perinuclear actin cap.

a Super resolution imaging of LoVo MET-KO cells stained with Phalloidin (red) and DAPI (blue) performed with X-Light V3 – DeepSIM microscope. Z-depth coding is reported as a scale of colors in the lower panel. Scale bar: 10 μm. b Focal adhesions (red) and actin cytoskeleton (green) visualization through Vinculin antibodies and Phalloidin staining in LoVo MET-KO and MET+ cells. Nuclei were stained with DAPI (blue). Pictures of basal and apical plane of cells are provided. Scale bar: 50 μm c Top and bottom view of a MET-KO cell stained with Phalloidin (green), Vinculin (red) and DAPI (blue) and visualized in 3D reconstruction. Yellow arrows point at the Actin Cap Associated Focal Adhesions (ACAFAs). Scale bar: 10 μm. The scale bar refers to the xy-plane. d Log2 fold change of filamin A (FLNA), actinin 4 (ACTN4), Rac Family Small GTPase 1 (RAC1) and Sad1 and UNC84 Domain Containing 1 (SUN1) expression in LoVo MET-KO cells compared to LoVo MET+ cells according to RNAseq data. ****Padj < 0.0001 **Padj < 0.01. e, f Pearson correlation between the expression of MET and FLNA (e) or ACTN4 (f) in colorectal cancer according to gene arrays data available on GEO. R and P-value are reported. g, h Non-muscle myosin IIA (green), actin cytoskeleton (Phalloidin – red) and nuclei (DAPI – blue) staining in MET-KO and MET+ cells. Pictures from different planes of section, as well as maximum intensity projection and 3D rendering are provided. Scale bar: 10 μm.