Figure 5.

GAS6-AS1 repressed E2F1-mediated transcription of GLUT1

(A) The potential binding site of E2F1 with GLUT1 promoter was shown, and the wild-type or mutant-type luciferase vectors were constructed. (B) Luciferase activity was assayed in A549 cells transfected with luciferase vectors (wild type or mutant type) and meantime co-transfected with expression plasmids (empty vectors, E2F1 expression plasmids, or GAS6-AS1 expression plasmids). (C) ChIP experiments of E2F1 (IgG as an internal control) were performed, and the co-precipitated DNA was subjected to PCR amplification with primers specific to GLUT1 promoter region. (D and E) The level of GLUT1 under ectopic expression of E2F1 or GAS6-AS1 was detected by qRT-PCR (D) and western blotting (E). (F and G) The correlation between E2F1 (F) and GAS6-AS1 (G) with GLUT1 in LUAD tissues is presented.