Skip to main content
. Author manuscript; available in PMC: 2010 Jan 22.
Published in final edited form as: J Immunol. 2008 May 1;180(9):5927–5934. doi: 10.4049/jimmunol.180.9.5927

FIGURE 2. E2F1-mediated transcriptional repression of Ifi202 is independent of p53 and pRb expression, but dependent on DNA-binding domain of E2F1.

FIGURE 2

(A) Sub-confluent cultures of wild type or p53-null MEFs were transfected with 202-luc-reporter plasmid (2.5 μg) and pRL-TK plasmid (0.5 μg) along with pCMV (2.5 μg, column 1) or increasing amounts (1 μg or 2 μg, columns 2 and 3, respectively) of pCMV-mE2F1 plasmid using a calcium phosphate transfection kit as described in methods. Cells were harvested after 44 h to assays for the firefly and Renilla luciferase activities as described in methods. Normalized relative luciferase activity is shown. Results from a representative experiment are shown.

(B) Sub-confluent cultures of p53-null MEFs were transfected with 202-luc-reporter plasmid (2.5 μg) and pRL-TK plasmid (0.5 μg) along with pCMV (2.5 μg, column 1) or increasing amounts (1 μg, 2 μg, or 3 μg, columns 2, 3, and 4, respectively) of pCMV-mE2F1 plasmid using a calcium phosphate transfection kit. Cells were harvested after 44 h to assays dual luciferase activities as described in (A) above. Normalized relative luciferase activity is shown. Results from a representative experiment are shown.

(C) Sub-confluent cultures of Rb-null MEFs were transfected with 202-luc-reporter plasmid (2.5 μg) and pRL-TK plasmid (0.5 μg) along with pCMV (2.5 μg, column 1) or increasing amounts (1 μg or 2 μg columns 2 and 3, respectively) of pCMV-mE2F1 plasmid using a calcium phosphate transfection kit. Cells were harvested after 44 h to assays dual luciferase activities as described in (A) above. Normalized relative luciferase activity is shown. Results from a representative experiment are shown.

(D) Sub-confluent cultures of p53-null MEFs were transfected with 202-luc-reporter plasmid (2.5 μg) and pRL-TK plasmid (0.5 μg) along with pCMV plasmid (2.5 μg, column 1) or plasmid (pCMV-mE2F1; 2.5 μg, column 2) encoding wild type mE2F1, or plasmid (2.5 μg) encoding various mutants of E2F1 [E2F1 (E132), column 3; E2F1 Δ206-220, column 4; E2F1 Δ1-88, column 5; and E2F1 (411/421) column 6] using a calcium phosphate transfection kit. Cells were harvested after 46 h to assays dual luciferase activities as described in (A) above. Normalized relative luciferase activity is shown. Results from a representative experiment are shown.