Figure 5

Comparison of expression levels of lymphangiogenesis-related factors among the clones. A) Each clone cultured was subjected to real-time RT-PCR for indicated factors. (n = 3 each, *p < 0.01 versus V1, **p < 0.001 versus V1, N.S. versus V1, # undetectable). B) Each clone cultured was subjected to RT-PCR for indicated growth factors. PCR primer sets are listed in additional file 4. cDNA templates from HMVEC and MRC5 were simultaneously amplified as positive controls. C) Each clone-derived tumor tissue was subjected to real-time RT-PCR for indicated human (h) growth factors using the human gene-specific primer sets listed in the additional file 4. (n = 3, *p < 0.001 versus V1, N.S. versus V1). D) Each clone-derived tumor tissue was subjected to RT-PCR for indicated human (h) growth factors using the human gene-specific primer sets listed in the additional file 3. cDNA templates from HMVEC and MRC5 were simultaneously amplified as positive controls. E) Tumor tissues derived from established clones (EBC1-V1, -V2, -P4, and -P15; n = 3 each) were subjected to real-time RT-PCR for human or mouse VEGF-C using the species-specific primer sets listed in the additional file 4. Plasmid templates inserting a part of human or mouse VEGF-C gene were prepared for standard curves as described in Methods. From the obtained data, the number of each template in the tumor-derived cDNA library was determined. The human VEGF-C mRNA level was expressed as a relative fold increase compared to that of the mouse VEGF-C level. Horizontal bar means average score.