Anti irf7

ChIP assays were performed with SimpleChIP Enzymatic Chromatin IP Kit (Cell Signaling Technology Japan, Tokyo, Japan) with anti-IRF1 (Santa Cruz Biotechnology), anti-IRF3(Santa Cruz Biotechnology), anti-IRF5 (Abcam), anti-IRF7 (Santa Cruz Biotechnology), anti-IRF8 (Santa Cruz Biotechnology), anti-IRF9 (Santa Cruz Biotechnology), normal goat IgG (Abcam) and normal rabbit IgG (Cell Signaling Technology) antibodies. ChIP signals were quantified by quantitative PCR analysis with a 7500 real-time PCR system (Applied Biosystems). Values obtained for immunoprecipitated samples (percent (%) input DNA) were normalized to values for respective normal IgG. The specific primer pairs for the Irf5 promoter region and P2rx4 promoter region, respectively, are described below.
Region 1: 5′-ATTTCTCAGGCCCTGTCTAAAGTG-3′ (forward),
5′-GGCACAGAGAGAGTTAGAGGAAGA-3′ (reverse)
Region 2: 5′-TATGGAGTCTTTCTGCACCCTGT-3′ (forward),
5′-TTCCAAGAACGAAGAGTCCCCTA-3′ (reverse)
ISRE-1: 5′-GCTGGCTCGTTTCAAGAATATT-3′ (forward),
5′-CGTACCCTGTAGCCGTCTATT-3′ (reverse)
ISRE-2: 5′-TCTACAGCCTGAAAGTCTATCATTG-3′ (forward),
5′-AAGGAATCTGAGAGGTACACACTG-3′ (reverse)
ISRE-3: 5′-GATAGGGAGAGGCTCGTTCA-3′ (forward),
5′-TAAAAGCTCGGGACCTGGAA-3′ (reverse)
ISRE-4: 5′-TACTGACCTGCCTCTTTTAAGGACA-3′ (forward),
5′-CGGAAAGAACTTTGAACCTTGAG-3′ (reverse)

Monoclonal antibodies (Abs), specific for cluster of differentiation (CD)1a, CD14, CD38, CD86, CD83, HLA-DR, CD40, IgG1, and IgG2a (BD Bioscience, San Diego, CA, USA), were directly conjugated to fluorescein isothiocyanate (FITC) or phycoerythrin (PE). To exclude dead cells from the analysis, Fixable Viability Dye eFluor®780 (FvDye) (eBioscience, San Diego, CA, USA) was used. For immunoblotting analysis, rabbit anti-STING (Cell Signaling, Danvers, MA, USA # 2775), anti-IRF3 (Santa Cruz, Santa Cruz, TX, USA # sc-9082), anti-IRF7 (Santa Cruz, # sc-9083), anti-STAT1 (BD Bioscience, # 610186), anti-phospho STAT1 (Cell Signaling Technology, Leiden, The Netherlands, # 7649), anti-STAT2 (BD Transduction Laboratories, # 610188), anti-phospho STAT2 (R&D Systems, Minneapolis, MN, USA, MAB2890), mouse anti-actin (Sigma-Aldrich, St. Louis, MO, USA #A0483), and horseradish peroxidase-conjugated secondary antibody anti mouse (Santa Cruz, # sc-2005) and anti rabbit (Santa Cruz, # sc-2004) were used. For phagocytosis and phagosomal acidification experiments, cytochalasin D 5 μM (Sigma-Aldrich, # C8723) and chloroquine 2 μM (Sigma-Aldrich, # C6628) were used.