JCS02483 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 - Figs S1, S3, S4. Immunohistochemical staining of perivascular SMA⁺ cells in Matrigel plugs with NG2 and desmin. We investigated perivascular SMA⁺ cells in Matrigel plugs using NG2 and desmin, which are reported to be markers of less mature pericytes. Each panel has four parts: NG2 or desmin staining (blue), SMA staining (red), PECAM1 staining (green) and the merged images, as indicated. Bars, 20 mm. Experimental conditions in Fig. S1 correspond to Fig. 1, Fig. S3 to Fig. 5 and Fig. S4 to Fig. 6.

• Supplemental Figure 2 - Fig. S2. Macrophage marker staining in VEGF-A-treated Matrigel plug. For further characterization of the cells in the VEGF-A-treated Matrigel plugs, cells were stained by F4/80, a macrophage marker, as well as with SMA and CD34. Bar, 20 mm.
• Supplemental Figure 3
• Supplemental Figure 4 -

• Movie 1 - Movies 1-4. Long-running video microscopic analysis of ESC-derived vascular cells. We observed ESC-derived VEGFR2⁺ cells treated with various growth factors in vitro by video microscopy for more than 12 hours. Video microscopy was started after a 24-hour incubation. Cells were cultured in the same conditions as shown in Fig. 1: (Movie 1) VEGF-A (30 ng/ml) and FGF-2 (10 ng/ml), (Movie 2) VEGF-A (30 ng/ml), (Movie 3) FGF-2 (10 ng/ml) and (Movie 4) PDGF-BB (20 ng/ml). Note that the cells that differentiated into SMA⁺ mural cells in the presence of VEGF-A and FGF-2 (Movie 1) are highly motile but remain close to endothelial sheets (see the cell indicated by an arrow as an example of such a mural cell).
• Movie 2
• Movie 3
• Movie 4