Figure 1: Induction of tumour-derived endothelial differentiation by Twist1.
From: Twist1 induces endothelial differentiation of tumour cells through the Jagged1-KLF4 axis
(a) Expression of CD31 and CD144 in Twist1-overexpressing OECM1 cells using western blot analysis. The middle panel showed the dot plots from flow cytometry analysis and the rightmost panel showed the percentage of cells expressing CD144 in Twist1-overexpressing OECM1 cells. (b) Co-expression of Twist1 and CD144 in Twist1-overexpressing OECM1 cells. Red: Twist1; green: CD144. (Left panels: images; right panel: the percentage of cells co-expressing Twist1 and CD144.) Scale bar, 50 μm. (c) 2.5-Dimensional (2.5D) tube-forming ability assays in Twist1-overexpressing OECM1 cells (left panels: images; right panel: pixels representing tube formation as described in Methods section (bar graph)). Scale bar, 50 μm. (d) DiI-AcLDL uptake assays in Twist1-overexpressing OECM1 cells. Red: DiI-AcLDL staining. The AcLDL staining photos were accompanied by 3D tube-formation ability photos (on top of Dil-AcLDL staining) for each clone. Scale bar, 50 μm. (e) Participation of GFP-labelled Twist1-overexpressing cells in tumour vessel formation. Green: GFP; red: CD144; blue: DAPI. Scale bar, 50 μm. (f) Co-expression of Twist1 and CD31 in HNC patient samples. Examples from two different patient samples are shown. Green: hCD31; red: Twist1; blue: DAPI. Scale bar, 50 μm. Data shown in panels a–c are mean±s.d. of three independent experiments. Asterisk (*) indicates statistical significance (P<0.05 by Student’s t-test) between experimental and control (cont.) clones. The OECM1 control clone was selected as the control group in panels a–c.
