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. 2016 Apr 20;7(21):31177–31190. doi: 10.18632/oncotarget.8870

Figure 3. ZNF224 DNA binding motif characterization.

Figure 3

A. DNA motif within ZNF224 enriched region from ChIP sequencing. B. Hair-pin loop structure containing 5′-CAGC-3′ sequence. C. Biotinylated oligo DNAs (5 μM) were fixed to streptavidin coated 96 well plates and purified FLAG-ZNF224 (50 ng) was loaded to each well. The bound FLAG-ZNF224 was detected with anti-FLAG antibody, and developed as described in the Methods section. Data are from at least three independent experiments (n=3). D. pGL3 vector containing (200 ng) 5′-CAGCTTC-3′ sequence with empty vector (E.V) or FLAG-ZNF224 (50, 100, and 200 ng) was transfected to MCF-7 cells for 24 h, and luciferase activity assay was performed. Luciferase activity was normalized to Renilla (100 ng) luciferase activity. The expression of FLAG-ZNF224 was detected with anti-FLAG antibody (inset). Tubulin was used as a loading control. Data are from at least three independent experiments (n=3).