class b cpg oligodeoxynucleotide odn 2006  (InvivoGen)

Journal: Developmental and comparative immunology

Article Title: Mechanisms of type I interferon production by chicken TLR21.

doi: 10.1016/j.dci.2023.105093

Figure Lengend Snippet: Fig. 2. CpG DNA drives a type-I interferon response in chicken macrophages (A) HD11 cells were stimulated with the indicated doses of type B CpG DNA (ODN2006) and transcription of IFNB and CCL4 was measured by qRT-PCR 6 h later. (B) HD11 cells were stimulated with the indicated PRR ligands (incubation with 5 μg ODN2006 or Poly(I:C), or transfection with 5 μg/ml htDNA, ctDNA or 2′3′-cGAMP) and transcription of the indicated genes was measured by qRT-PCR 6 h later.

Article Snippet: The Class B CpG ODN2006 (ODN 7909, PF_3512676, sequence: 5′-tcgtcgttttgtcgttttgtcgtt-3′, Invivogen) was diluted in sterile endotoxin-free water.

Techniques: Quantitative RT-PCR, Incubation, Transfection

Journal: Developmental and comparative immunology

Article Title: Mechanisms of type I interferon production by chicken TLR21.

doi: 10.1016/j.dci.2023.105093

Figure Lengend Snippet: Fig. 3. TLR21 expression is enhanced by the synergistic activity of IFN-I and CpG HD11 cells were preincubated with chicken interferon alpha (50 ng/ml for 16 h) before being stimulated with type 5 μg B CpG DNA (ODN2006) for the indicated times. Transcription of IFNB and CCL4 was subsequently measured by qRT-PCR.

Article Snippet: The Class B CpG ODN2006 (ODN 7909, PF_3512676, sequence: 5′-tcgtcgttttgtcgttttgtcgtt-3′, Invivogen) was diluted in sterile endotoxin-free water.

Techniques: Expressing, Activity Assay, Quantitative RT-PCR

Journal: Developmental and comparative immunology

Article Title: Mechanisms of type I interferon production by chicken TLR21.

doi: 10.1016/j.dci.2023.105093

Figure Lengend Snippet: Fig. 5. CpG-driven interferon production is independent of TBK1. HD11 cells were stimulated with ODN2006, or by transfection with htDNA or ctDNA, in the presence or absence of (A) BX795 or (B) BAY-11-7082. Transcription of the indicated genes was measured by qRT-PCR 6 h later. NS- non significant, **p < 0.01, ***p < 0.001, n ≥3.

Article Snippet: The Class B CpG ODN2006 (ODN 7909, PF_3512676, sequence: 5′-tcgtcgttttgtcgttttgtcgtt-3′, Invivogen) was diluted in sterile endotoxin-free water.

Techniques: Transfection, Quantitative RT-PCR

Journal: Developmental and comparative immunology

Article Title: Mechanisms of type I interferon production by chicken TLR21.

doi: 10.1016/j.dci.2023.105093

Figure Lengend Snippet: Fig. 6. TLR21 is the primary CpG DNA sensor and signals via IRF7 in chickens. (A) TLR21 knockout and (B) IRF7 knockout HD11 cells were generated by CRISPR/Cas9 editing and sequenced to define indels targeted by the gRNAs (yellow) including PAM sequences (red). (C) WT, TLR21 KO and IRF7 KO HD11 cells were stimulated with 5 μg ODN2006, or by transfection with 5 μg/ml htDNA or ctDNA and transcription of the indicated genes was measured by qR T-PCR 6 h later. (D) WT, TLR21 KO and IRF7 KO HD11 cells were stimulated with 5 μg ODN2006, or by transfection with 5 μg/ml htDNA or ctDNA and cell viability measured 24 h later by flow cytometry. NS- non significant, **p < 0.01, ***p < 0.001, n ≥3. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The Class B CpG ODN2006 (ODN 7909, PF_3512676, sequence: 5′-tcgtcgttttgtcgttttgtcgtt-3′, Invivogen) was diluted in sterile endotoxin-free water.

Techniques: Knock-Out, Generated, CRISPR, Transfection, Flow Cytometry