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 Jul 4;10:1426. doi: 10.3389/fmicb.2019.01426

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

Copyright © 2019 Hurst, Aswad, Karamitros, Katzourakis, Smith and Magiorkinis.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

PMC Copyright notice

IFI16 binds to oligos from HIV-1 and HERV-K. Pre-cleared Jurkat cell lysates were incubated with biotin-labeled oligos, as described in the section “Materials and Methods.” The pulldowns were analyzed by SDS-PAGE and western blotting first for IFI16, followed by reprobing the same blot for cGAS. (A) The ssDNA 1, 2 and 3 oligos (Jakobsen et al., 2013) were examined for their interaction with IFI16 (top panel) and cGAS (bottom panel). The lanes were loaded with input lysate or pulldowns with oligos as indicated: (1) input lysate, (2) ssDNA1, (3) ssDNA2, (4) ssDNA3, (5) no oligo control and (6) repeat of the input lysate. (B) A HK2 oligo from the consensus sequence, as well as the HK113 sequence, were analyzed for interaction with IFI16 (top panel) and cGAS (bottom panel). The lanes were loaded with input lysate or pulldowns with oligos as indicated: (1) input lysate, (2) GC59, (3) ssDNA1, (4) HK2 env, (5) HK2 env mut, (6) HK2 env scrambled 2, (7) HK113 LTR, (8) HK113 env and (9) a positive input lysate from a previous experiment as a blotting control. (C) In this competition assay, the biotinylated ssDNA1 and HK113 LTR oligos were used in pulldowns with and without an untagged version of the same oligo. The lanes were loaded as follows: (1) input lysate, (2) HK113 LTR biotinylated oligo only, (3) HK113 LTR biotinylated and unlabeled oligos, (4) ssDNA1 biotinylated oligo only, (5) ssDNA1 biotinylated and unlabeled oligos, (6) empty lane and (7) repeat of the input lysate. Densitometry was performed on the IFI16 blots by measuring the area under the curve (AUC) with Image J.