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. 1999 Mar 16;96(6):3071–3076. doi: 10.1073/pnas.96.6.3071

Figure 4.

Figure 4

VEGFR binding specificity of ORFV2-VEGF. Soluble fusion proteins consisting of the extracellular domain of VEGFRs and the Fc portion of human IgG1 were used to assess the receptor-binding specificity of ORFV2-VEGF. Biosynthetically labeled conditioned medium derived from 293EBNA cells transfected with ORFV2-VEGF, mouse VEGF164, human VEGF165, human VEGF-DΔNΔC, human VEGF-C156S, or vector alone were immunoprecipitated with VEGFR-1-Ig, VEGFR-2-Ig, VEGFR3-Ig, or NP-1 fusion proteins bound to protein A. Labeled growth factors immunoprecipitated with VEGFR-1-Ig, VEGFR-2-Ig, VEGFR3-Ig fusion proteins and M2-gel (anti-Flag) (A) or NP-1-Ig and VEGFR-1-Ig (B) were eluted from washed beads with SDS/PAGE sample buffer and resolved by using SDS/PAGE (15%). Note that only the ORFV2-VEGF and VEGF-DΔNΔC are Flag-tagged. Dried gels were visualized by using PhosphorImager analysis. The fusion protein used for each precipitation is listed above each panel.