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. 2017 Apr 3;7:45815. doi: 10.1038/srep45815

Figure 4. H19 G-quadruplex suppresses transcription.

Figure 4

(A) Schematic diagram of the H19 expression vectors used in this study. The full length H19 gene (WT-H19), G-quadruplex sequence-truncated H19 gene (∆G4-H19), or H19 gene with Mut2 or Mut3 G-quadruplex sequence (Mut2-G4-H19, Mut3-G4-H19) were driven by the EF1α promoter. (B) qPCR analysis of H19 RNA levels in 293T cells transfected with the indicated plasmids. Values are normalized against G3PDH. (C) PCR analysis for transfection efficiency of the plasmids in (B) using a primer pair specific to the vector backbone. (D) Schematic diagram of the luciferase reporter plasmids. H19 G-quadruplex sequence was inserted just upstream of the luciferase gene (Luc). Luc or G-quadruplex-Luc (G4-Luc) was driven by the H19 promoter (the region between −840 to +14). (E,F) 293 T cells (E) or EpH4 cells (F) were transfected with the luciferase reporter plasmids together with the control RL-SV40 plasmid, and the luciferase activities measured. Values are normalized against the activity of co-transfected Renilla luciferase. (B,E,F) mean ± s.d. from three experiments; *P < 0.05, ***P < 0.005 analyzed by Dunnett’s multiple-comparison test.