FIGURE 4.

E2F1-dependent expression of Bim in cancer cells. A, Bim promoter constructs of indicated fragment size were cloned in pGL3-basic vector and E2F1-binding sites were mutated as shown. B and C, LNCaP cells were transfected with −1.6, −2.6, or −5.1-kb Bim promoter luciferase (luc) constructs (B) or −5.1-kb fragment and its mutants (C) as described under “Experimental Procedures” or empty vector together with Renilla luciferase construct. D, MDA-MB231 cells were infected with control or E2F1 shRNA lentiviral particles at a multiplicity of infection of 5. Stable cells were subjected to Western blotting to detect indicated proteins. Actin serves as loading control. E, NHP9 normal cells and LNCaP cancer cells were transfected with −5.1-kb Bim promoter construct. Luciferase activities in B, C, and E were determined at 24 h after transfection. RLU, relative luciferase units. Data are mean ± S.D., n = 3; *, p < 0.01.