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. 2021 Aug 2;11:15655. doi: 10.1038/s41598-021-95165-6

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Segment C of Exon3 may contain a G-quadruplex. (a) Wild-type and mutant oligonucleotides (red font) of Segment C together with Pu₂₂ of VEGF with possible G4 forming sequences (underlined) are shown. (b, c) Assessing the presence of a G4 structure in Segment C using Thioflavin T (ThT). (b) Fluorescence emission of ThT in the presence of selected nucleic acid sequences (c) bar graph of the change in ThT fluorescence in the presence of selected nucleic acid sequences. (d) CD spectra of Segment C oligonucleotides at 5 °C. (e) Thermal denaturation profile of Segment C oligonucleotides obtained from the change in UV–Vis absorbance at 295 nm between 15 to 95 °C. (f) MCF7 cells were transiently transfected with pGL3 bearing none (Basic-Luc), the CMV promoter without (CMV-Pr), or with the 3′-end genetically fused Segment C (CMV-Pr-C), Segment D (CMV-Pr-D), or Segment C bearing the G4 sequence deletion (CMV-Pr-CΔ_G4). Reporter vectors drive the expression of the Firefly Luciferase cDNA expression as the reporter enzyme. The transfection efficiency was monitored by the co-expression of pCMV-RL that drives the expression of the Renilla Luciferase cDNA. 24 h after transfections, cellular extracts were subjected to dual-luciferase assays. Shown are the mean ± SE of three independent experiments performed in triplicate. Firefly/Renilla Luciferase activities are presented as fold change (log₁₀) compared to pGL3-Basic, which is set to one. Asterisk (*) indicates significant differences from the Basic-Luc control.