Mammalian Target of Rapamycin Inhibitors Induce Tumor Cell Apoptosis In Vivo Primarily by Inhibiting VEGF Expression and Angiogenesis
Figure 3
Characterization of exogenous VEGF. (a) Cartoon of plasmids used to generate exogenous VEGF. The p27 IRES was cloned upstream of the VEGF ORF. (b) Stably transfected isogenic HS Sultan cell lines or HS Sultan parental cell line were treated with indicated concentration of rapamycin for 48 hours. ELISA was used to determine the VEGF levels/mg of protein in cell supernatants in triplicate (data are presented as mean ± SD). (c) The levels of FLAG-tagged VEGF were measured in cellular supernatants (in triplicate) using anti-FLAG ELISA and anti-VEGF secondary antibody (data are presented as mean ± SD).
