Fig. 3.
TIP-1 knockdown impairs production of angiogenic factors within D54 cells. (A) Matrigel-plug assays. A half million D54 cells with or without TIP-1 knockdown were mixed with matrigel for subcutaneous implantation in nude mice. FITC-dextran was injected into the tail vein on day 8 after the matrigel plug implantation. Tumors were resected for examination with a confocal fluorescence microscope. Shown are representative images and quantitative measurement (ImageJ) of the FITC signals from at least 12 microscopic fields. * p<0.05 compared to the shRNA control. (B) Angiogenic activity of pre-conditioned medium from monolayer culture of D54 cells was analyzed with a Boyden chamber-based HUVEC migration assay. Shown are the means ± SDs of three experiments in which HUVECs migrated through the porous chambers. (C) HUVEC tubule formation assay using pre-conditioned medium from monolayer culture of D54 cells. Shown are the quantitative measurements of capillary connections with more than 3 branches. Bars represent the means ± SDs of three independent experiments. * p<0.05 compared to the shRNA control by the Student’s t-test.