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. 2015 Mar 14;6(13):11252–11263. doi: 10.18632/oncotarget.3594

Figure 2. Over-expression of E2F1 reduces SIRT6 transcription through direct binding to SIRT6 promoter.

Figure 2

(A) E2F1 over-expression reduced SIRT6 protein and its histone deacetylase activity. (B) Reduction of SIRT6 mRNA levels at 12 hours and 24 hours post transfection as detected by qRT-PCR in pcDNA-E2F1 transfected cells (E2F1) versus vector transfected cells (V). (C) Dual reporter luciferase assay comparing SIRT6 promoter activities with different combinations of pcDNA3 constructs [vector, E2F1, E2F1(138)] and SIRT6 promoter PGL3 reporter constructs (SIRT6 and SIRT6 with mutated E2F1 binding site, mSIRT6). The sequence of the E2F1 binding site on the proximal SIRT6 promoter and mutated sites on this sequence were shown above (D) Compared to vector transfected cells (V), E2F1 over-expression (E2F1) enhanced binding of E2F1 to the SIRT6 promoter on the ChIP assays. Representative gel pictures were shown above and the intensities of E2F1 binding were quantitated as bar graphs below. Each bar represents the mean and SEM of 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001.