The TEAD4216 Isoform Inhibits VEGF Expression Figure 6. The TEAD4216 isoform can reduce endogenous VEGF165 protein. Transient plasmid transfection or stable lentiviral mediated introduction of TEAD4216 into human cells results in reduction of native VEGF protein. Solid bars represent VEGF165 levels, quantified by ELISA, within conditioned media collected 48 hours after transfection of pcDNA plasmid vector containing the TEAD4216 isoform into (A) 293T cells, (B) ARPE-19 retinal pigment epithelial (RPE) cells in culture (n = 4), (p,0.05). (C) Lentiviral (LV) expression of the TEAD4216 isoform in human D407 RPE cells can inhibit endogenous VEGF production. Solid bars represent VEGF165 levels, quantified by ELISA, within conditioned media collected 48 hours after transduced RPE cells were FAC sorted and plated (n = 3),(p,0.02). doi:10.1371/journal.pone.0031260.g006 Figure 7. Lentiviral (LV) mediated expression of the TEAD4216 isoform in 293T cells inhibits cell proliferation. A Cyquant cell proliferation assay determining the DNA content of cells indicate that 4 days after initial seeding of equal cell numbers the control untransduced cells proliferate at a faster rate than the LV-TEAD4216 transduced cells (n = 8, p,0.05). doi:10.1371/journal.pone.0031260.g007 PLoS ONE | www.plosone.org 5 June 2012 | Volume 7 | Issue 6 | e31260