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2 protocols using poly 1 c hmw tlr 3 ligand

1

Reagents for Molecular and Cell Biology

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Cyanidin chloride (A18; CAS 528-58-5) was obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Pam2CSK4 (TLR2/TLR6 ligand) and poly(I:C)-HMW (TLR 3 ligand) were purchased from InvivoGen (San Diego, CA, USA). The mouse monoclonal antibody anti-gF of RSV was obtained from US Biological Life Sciences (Salem, MA, USA). The rabbit polyclonal antibody anti-Spike of SARS-CoV-2 was obtained from Sigma-Aldrich (St. Louis, MO, USA). Secondary goat anti-mouse FluoroLinkTM CyTM3 and goat anti-rabbit FluoroLinkTMCyTM2 antibodies were purchased from GE Healthcare (Chicago, IL, USA). DAPI was purchased from Sigma-Aldrich (St. Louis, MO, USA).
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2

Toll-Like Receptor Signaling Assays

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LPS (TLR4 ligand) from Escherichia coli serotype 055:B5, (−)-N6-(2-Phenylisopropyl) adenosine (R-PIA), dibutyryl cAMP (dbcAMP) and forskolin were obtained from Sigma. Pam2CSK4 (TLR2/TLR6 ligand), poly(I:C)-HMW (TLR 3 ligand), imiquimod (TLR7 ligand) and CpG ODN 2395 (TLR9 ligand) were purchased from InvivoGen. Resiquimod (TLR7/8 ligand) was kindly provided by Dr. Marianela Candolfi (INBIOMED-UBA-CONICET). Imiquimod was dissolved in water, according to the manufacturer's instructions. The rabbit monoclonal anti-Phospho-CREB (Ser133) (87G3) and anti-CREB (48H2) were purchased from Cell Signaling. KT5720 was obtained from Tocris Bioscience. The mouse monoclonal antibody anti-gF of RSV was obtained from US Biological Life Sciences. Secondary goat anti-mouse FluoroLinkTM CyTM3 antibodies were purchased from GE Healthcare. The peroxidase-conjugated goat anti-rabbit antibodies and Dapi for nucleic acid staining were obtained from Sigma.
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