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. 2011 Oct 18;6(10):e25826. doi: 10.1371/journal.pone.0025826

Figure 4. Sema4D treatment of HUVEC induces NF-κB-dependent production of IL-8, which is necessary to promote angiogenesis.

Figure 4

A. ELISA for production of IL-8 in HUVEC treated with the indicated concentrations of Sema4D or 400 ng/ml Sema4D with BAY11-7085 (BAY). Results are expressed as averages in pg/ml. Error bars represent the standard deviation for three independent experiments. B. Cell migration assay for HUVEC migrating towards BSA or Sema4D in the presence of IgG control (IgG) or the indicated concentrations of anti-IL-8 blocking antibody. The results are quantified in the bar graph below as the pixel intensity of the scanned migration assay membrane. Error bars represent the standard deviation from six wells (*, p<0.05). C. HUVEC were plated on reconstituted basement membrane material in control media (BSA) or media containing Sema4D, with IgG control (IgG) or the indicated concentrations of IL-8 blocking antibody and examined for formation of capillary tubes. Representative photographs are shown. Quantification of the results observed in the tubulogenesis assay are shown in the bar graph below (error bars represent the standard deviation from three independent experiments; *, p<0.05).