1-IGFBP7. Moreover, many biological roles of pcDNA3.1-IGFBP7 remain to be elucidated. Acknowledgements We thank Ming jian Yang for technique guidance, and Hoi Lun Lau for editing the manuscript. This project was supported by the National Science Fund Program from the National Natural Science Foundation of China (No. 30700717). Electronic supplementary material Additional file 1: pcDNA3.1-IGFBP7 plasmid checked by restriction enzyme analysis, and transfection with Effectene authenticated by immunofluorescence. Restriction enzyme analysis of pcDNA3.1-IGFBP7 plasmid by EcoR I BKM120 and Bgl II manifested that the obtained plasmid was the objective one with predicted length. Plasmid transfection with Effectene was successful, authenticated

by immunofluorescence. (PDF 75 KB) Additional file 2: Effect of pcDNA3.1-IGFBP7 plasmid on IGFBP7 expression in vitro. Higher concentration of pcDNA3.1-IGFBP7 plasmid led to higher IGFBP7 mRNA and protein expression in B16-F10 melanoma cells, detected by RT-PCR and western blot. pcDNA3.1-IGFBP7 transfection led to reduction of B16-F10 cells viability, check details determined by the Cell Counting Kit-8. (PDF 256 KB) Additional file 3: Effect of different plasmids on tumor cell apoptosis rate

detected by flow cytometry and laser scanning confocal microscopy. Apoptosis rate detected by flow cytometry of B16 melanoma resulted in an obvious increase in pcDNA3.1-IGFBP7 group than those in pcDNA3.1-CONTROL and B16 groups, consistent with laser confocal display of tumor sections of the three groups, suggested significant effects of in-vitro and in-vivo pcDNA3.1-IGFBP7 Chlormezanone transfection on B16 apoptosis. (PDF 444 KB) Additional file 4: In-vivo anti-tumor effect of pcDNA3.1-IGFBP7 plasmid. Survival curves and tumor volumes showed different effects of the three groups. pcDNA3.1-IGFBP7 group has a significantly higher survival rate and smaller tumor size, compared to pcDNA3.1-CONTROL and B16-F10 groups. (PDF 127 KB) References 1. Zheng H, Gao L, Feng Y, Yuan L, Zhao H, Cornelius LA: Down-regulation

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