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

. 2021 Mar 31;11:7243. doi: 10.1038/s41598-021-86644-x

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

© 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/.

PMC Copyright notice

Cytosolic DNA Levels Do Not Contribute to the STING Dysfunction in Human Cancer Cells. (A) TRAMP-C2, THP-1, DU145, A549, HeLa, and HCT116 cells were stained for dsDNA or RNA:DNA hybrids recognized by the S9.6 antibody (red) in presence of DAPI (blue). (B) Cytosolic dsDNA (left panel) and RNA:DNA hybrids (right panel) in A549 cell lysates were immunoprecipitated with dsDNA-specific or S9.6 antibodies. Precipitates were immunoblotted using a cGAS-specific antibody. A portion of the lysates was treated with 20 units DNase or RNaseH prior to elution. The shown blots are representative of 3 independent experiments. (C) A549 or HCT116 cells were transduced with a retrovirus encoding TREX-1 or control vector and were stained for dsDNA (red) in presence of DAPI (blue). Bar graph shows quantification of the dsDNA MFI ± SD in TREX1- (white column) and control- (black column) transduced A549 and HCT116 cells. Values were normalized to the MFI in control cells. See also Figure S3. (D) RNASEH1 (RNH)- or control-transduced A549 and HCT116 cells were stained for RNA:DNA hybrids (red) in the presence of DAPI (blue). Bar graph shows quantification of the RNA:DNA hybrid MFI ± SD in RNASEH1 (RNH)- (white column) and control- (black column) transduced A549 and HCT116 cells. Values were normalized to the MFI in control cells. See also Figure S3. (E) IFNA (black column), IFNB (white column) and CXCL10 (grey column) transcript levels in TREX1- or RNASEH1 (RNH)-transduced A549 and HCT116 cells were determined by real-time PCR. Transcript levels were normalized to transcript levels in control-transduced cells. (F) TREX1- and RNASEH1 (RNH)-transduced A549 and HCT116 cells were treated with PBS (Ctrl) or 2 μg/ml cGAMP (cGa) for 4 h. IFNA (black column) and IFNB (white column) transcript levels were measured by real-time PCR and normalized to control cells. Scale bar denotes 10 μm in all images. All data are presented as mean ± SD of 3 independent experiments.