Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2019 Jan 30;47(7):3365–3382. doi: 10.1093/nar/gkz041

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

PMC Copyright notice

Figure 4. — Extracellular release of VEGF-A is regulated by TRF2 through the control of SULF2 expression. (A) Gene expression of SULF2 was evaluated by qPCR in HCT116 cells silenced (shTRF2) or overexpressing (pBabe-TRF2) TRF2 and in their control counterparts (shScramble and pBabe). Results are expressed as fold change of mRNA levels in silenced/overexpressing cells over their controls, after β-actin normalization. (B) IF analysis of SULF2 expression in cells silenced or overexpressing TRF2 and their controls. Representative images acquired at 63x magnification are shown. (C) Expression of TRF2 (left panel) and SULF2 (right panel) was evaluated by qPCR in HCT116 cells overexpressing the wild-type (pBabe TRF2) or the mutated forms of TRF2 (pBabe TRF2ΔM and pBabe TRF2ΔBΔM) and their control counterpart (pBabe). Results are expressed as fold change of mRNA levels in overexpressing cells over their controls, after β-actin normalization. (D) Schematic representation of putative TRF2-binding sites located upstream the transcription starting site (TSS) of SULF2. (E) Real-time qPCR analysis of TRF2-chromatin immunoprecipitates. DNA regions containing (Chr.2 sub-telomeric region) or not (RPLP0) TTAGGG sequences were used as positive and negative control, respectively. (F) pGL3-promoter vector (pGL3 empty) and the vector containing the wild-type or mutant form of the distal regulatory element were co-transfected with a renilla vector (pRL-TK) in control (pBabe) and TRF2 silenced (shTRF2) HCT116 cells and luciferase activity was assayed. Firefly luciferase signal was normalized for the renilla signal to derive the relative luciferase activity. Results are expressed as fold change over the activity measured in the cells transfected with the control vector. The histograms show the mean ± SE of at least three independent experiments performed in triplicate (*P < 0.1, **P < 0.01, ***P < 0.001; Student's t-test).