DNA-PK is a DNA sensor for IRF-3-dependent innate immunity
- Imperial College London, United Kingdom
- University of Cambridge, United Kingdom
Figures
Figure 1
DNA-PK binds DNA in the cytoplasm.
(A) Biotinylated DNA or biotinylated pam-3-cys were transfected into HEK 293T cells and 30 min later DNA was affinity purified from the cytoplasmic fraction before analysis of DNA/protein complexes …
Figure 2
The innate immune response to DNA requires DNA-PK in fibroblasts.
(A) ISD DNA of different lengths was transfected into MEFs and the transcription of Cxcl10 was assayed by qPCR 6 hr later. (B) Double stranded oligonucleotides (bio-ISD), concatenated ISD DNA …
Figure 3
IRF-3 and TBK1 are required for the stimulation of multiple genes in response to DNA.
(A) Primary wild type or Irf3−/− MEFs were transfected with DNA and the level of induction of Ifnb, Il6, Cxcl10 and Isg54 mRNAs were measured by qRT-PCR 6 hr later. (B) Immortalised wild type or Tbk1…
Figure 4
DNA-PK activates IRF-3-dependent, NF-κB–independent signalling.
(A) The localisation of endogenous IRF-3 was analysed by immunofluorescence 1 hr after transfection of primary wild type or Prkdc−/− MEFs with DNA or poly (I:C) (left panels) and quantified by …
Figure 5
DNA-PKcs kinase activity is dispensable for the innate immune response to DNA.
Primary fibroblasts from Balb/c or PrkdcSCID mice were transfected with DNA or poly(I:C) or infected with MVA or NDV and the level of induction of (A) Ifnb, (B) Cxcl10, or (C) Il-6 was measured 6 hr …
Figure 6
DNA-PK and TBK1 localise to sites of VACV DNA replication in infected cells.
HeLa cells were untreated or infected with MVA (m.o.i. = 5) for 6 hr. Cells were then fixed and stained with antibodies against (A) Ku80 or DNA-PKcs, and (B) TBK1 or IRF-3. Cytoplasmic viral …
Figure 7
DNA-PK contributes to the IRF-3-dependent innate immune response to MVA.
(A) Ifnb and Cxcl10 transcription was measured 6 hr following MVA infection of primary WT and Irf3−/− fibroblasts at an m.o.i. of 5. (B) Wild type and Prkdc−/− MEFs were infected with MVA or NDV and …
Figure 8
DNA-PKcs contributes to the innate immune response to MVA, HSV-1 and DNA in vivo.
(A) Groups of five Prkdc+/+ and Prkdc−/− mice were injected intradermally into the ear pinna with cationic lipids complexed with 1 μg DNA or poly(I:C). RNA was extracted from the tissue 12 hr later …
