Figure 3

Knockdown of URG4/URGCP reduces the angiogenic capacity of HCC cells in vitro. A. Western blotting analysis of URG4/URGCP protein expression in URG4/URGCP-shRNA-transduced QGY7703 and Hep3B cell lines (shown as URG4/URGCP-RNAi) and the corresponding vector control cells; α-Tubulin was used as a loading control. The numbers represent the relative expression of each protein compared to the respective control cells. B. Representative images (left) and quantification (right) of tube-like structures formed by HUVECs on Matrigel-coated plates when cultured in conditioned medium (CM) derived from the indicated cells. C. Representative images (left) and quantification (right) of the number of migrated HUVEC cells when incubated in CM derived from the indicated cells in the Transwell migration assay. D. Representative images (left) and quantification (right) of neovessels formed in the CAM assay when stimulated by CM derived from the indicated cells. E. Quantitative real-time PCR analysis of VEGFC mRNA expression in the indicated cells. Transcript levels were normalized to GAPDH and expressed relative to the respective control cells. F. ELISA of VEGFC protein expression in the indicated cell supernatants. Data is mean ± SD of three independent experiments; * P < 0.05.