Figure 2.

Effects of recombinant canstatin on the expression of vascular endothelial growth factor (VEGF) family proteins in CoCl₂‐treated squamous cell carcinoma (SCC)‐VII cells. (A) SCC‐VII cells were treated with different concentrations of recombinant canstatin (0, 0.5, 40 μg/mL) in the presence of 100 μmol/L CoCl₂, and incubated for 24 h. cDNAs were generated from DNase I‐treated total RNA, and PCR was performed with specific primers for vascular endothelial growth factor (VEGF)‐A, ‐B, ‐C, and β‐actin. (B) The PCR products from three independent experiments in (A) were quantified and are represented as a bar diagram. The transcript levels of VEGF‐A, ‐B, and ‐C mRNA in the control (recombinant canstatin‐ and CoCl₂‐untreated cells) were established as 100%. (C) Protein levels of VEGF‐A and ‐C in the intracellular fraction were determined using western blot analysis with anti‐VEGF‐A and anti‐VEGF‐C antibodies. (D) The amounts of VEGF‐A and ‐C obtained in three independent experiments of (C) were quantified and are represented as a bar diagram. The levels of VEGF‐A and ‐C in the control were established as 100%. Data are presented as mean ± SD of three independent experiments (*P < 0.05, **P < 0.01, ***P < 0.001).