Podoplanin-Fc inhibits lymphatic vessel formation in mouse EBs in vitro. (A) Lymphatic and blood vessel-like structures were visualized by immunofluorescent staining for the lymphatic marker LYVE-1 (green) and the pan-endothelial marker CD31 (red), in mouse EBs exposed to human VEGF-A (20 ng/mL), human VEGF-C (200 ng/mL), and podoplanin-Fc or human IgG (1μM) from days 14 to 22 after EB formation. Bars represent 100 μm. (B) LYVE-1 and CD31 double-positive lymphatic vessel-like structures and (C) CD31 positive blood vessel-like structures per EB were quantified. The number of individual structures is depicted as a boxplot, representing the median, minimum, and maximum data values and upper and lower quartiles. * P = .013 (n = 18).