Fig. 5: E2F1 directly represses the expression of miR-1205.

a RT-qPCR analysis of the relative miR-1205 expression in Hep-2 and KB-3-1 cells expresses Flag and Flag-E2F1. b Schematic representation of the miR-1205 locus on chromosome 8. The red squares indicated seven putative E2F1-binding sites, while the yellow arrow indicated transcription start sites. Three blue bars indicated truncated promoter fragments, respectively. Three mutations of binding sites were Mut1, Mut2, and Mut3. The contents of the amplification represent different mutation sites, and the red bases stand for the mutated bases. The F1, F2, R1, and R2 arrows represented different ChIP-PCR primers of BS2. c, d HEK293T cells were transfected with a luciferase reporter construct containing the indicated genomic regions and mutants with or without Flag-E2F1 plasmids. Relative luciferase activities were measured 24 h after transfection. The results are depicted as the ratio of Firefly luciferase and Renilla luciferase activity. e PCR gel showing E2F1 was enriched in the binding site 2 (BS2) of the putative miR-1205 promoter by ChIP assay. Data are presented as mean ± s.d. Student’s t-test was used for statistical analysis; *P < 0.05, **P < 0.01.