Figure 2. tGLI1 is associated with increased VEGF-A gene expression in human breast cancer and medulloblastoma cells.
A: Isogenic MDA-MB-231-GLI1 and MDA-MB-231-tGLI1 cell lines express GLI1 and tGLI1, respectively, at levels similar to those in natural breast cancer cells. The encoded tGLI1 protein (146 kD) was distinguished from the GLI1 protein (150 kD) using prolonged 5.5% SDS-PAGE followed by WB with an antibody that recognizes both GLI1 and tGLI1 isoforms. Top, WB. Bottom, RT-PCR.
B. VEGF-A gene is expressed at a higher level in tGLI1-expressing MDA-MB-231 cells than those with control vector and GLI1. Top, RT-PCR. Bottom, WB.
C: Expression of VEGF-A in breast cancer and medulloblastoma cells transiently transfected to express GLI1 or tGLI1, as shown by RT-PCR.
D: Human breast cancer cell lines with low tGLI1 levels expressed lower levels of VEGF-A than those with high levels of tGLI1, in which levels of tGLI1 were determined previously in Fig. 1B. WB was conducted.