Fig. 3

Maximal VEGFR2 activation requires Sdc2 HS chains. a–f Primary mouse ECs from indicated genotypes were serum-starved for 12 h and then stimulated for 5 and 15 min. Activation of VEGFR2 (pVEGFR2) was assessed after stimulation with VEGFA₁₆₅ (50 ng/ml) in Sdc2^−/− ECs (a representative picture, b quantification) or Sdc4^−/− ECs (c representative picture, d quantification). FGFR1 signaling in Sdc2^−/− EC was assessed by stimulation with FGF2 (20 ng/ml) and evaluation of ERK activation (e representative picture, f quantification) (n = 3–4 for VEGFA₁₆₅, n = 3 for FGF2). g, h, Sdc2^−/− ECs were transduced with adenovirus expressing the indicated construct for 16 h (MOI = 1–2), starved for 12 h followed by stimulation with VEGFA₁₆₅ (50 ng/ml) for 5 min. Rescue of VEGFR2 activation for indicated construct is shown (g representative picture, h quantification) (n = 3–6). Errors bars represent SEM. (N.S. not significant, ^*P < 0.05, ^**P < 0.01, ^***P < 0.001, N.S. not significant, by one-way Anova with Bonferroni’s multiple comparison test)