Figure 4.

Effects of podoplanin knockdown or of platelets lacking CLEC-2 on LEC migration. A) LEC settled onto a Transwell filter were treated with or without platelets (10⁸), with or without siRNA targeting podoplanin, with or without treatment with VEGF-C (300 ng/ml). The percentage of the LEC which had transmigrated after 24 h was analyzed. Data are mean ± SEM from three experiments. ANOVA showed significant effects of VEGF-C, siRNA treatment and platelet treatment (p < 0.01 in each case). **=p < 0.01 compared to nonspecific siRNA by Dunnett’s test, for VEGF-C-treated LEC. B) LEC settled onto a Transwell filter were treated with or without siRNA targeting podoplanin, with or without treatment with VEGF-A (30 ng/ml). The percentage of the LEC which had transmigrated after 24 h was analyzed. Data are mean ± SEM from at least three experiments. ANOVA showed significant effects of VEGF-A and of siRNA treatment (p < 0.01 in each case). **=p < 0.01 compared to nonspecific siRNA by Dunnett’s test, for VEGF-A-treated LEC. C) Platelets (10⁸) were added to LEC which had been settled onto a Transwell filter in the presence of VEGF-C (300 ng/ml). The platelets were from Clec1bfl/fl (CLEC-2⁺) or Clec1bfl/flPF4-Cre (CLEC-2⁻) mice. Wells without platelets received 100μl Ca2+/Mg2+-free sterile PBS. The percentage of the LEC which had transmigrated after 24h was analyzed. Data are mean ± SEM from at least four experiments. ANOVA showed that platelets had a significant effect (p < 0.01). ** = p < 0.01 by Bonferroni post-test.