Cytokine quantities for IL-6, TNF-a, IL-10, CXCL1, IL-12p40, and IFN-g in the lung tissue were determined by the mouse proinflammatory 7-plex tissue culture kit (Meso Scale Discovery, cat K15012B-1) using 25ul of homogenized lung tissue according to manufacturer’s instructions. The concentration of IFN-α and IFN-β from mouse cell supernatant and mouse lung tissue was measured with VeriKine mouse IFN-α ELISA kit (PBL, cat 42400) and VeriKine mouse IFN-β ELISA kit (PBL, cat 42400), respectively.
The mRNA level of TNF-α, IFN-β, IL-6, and IFN-α in GM-DCs were tested with SYBR green method as described above. Primers used for the cytokine test were the following:
TNF-α-F, 5'-GGCAGGTCTACTTTGGAGTCATTG-3’, TNF-α-R, 5'-ACATTCGAGGCTCCAGTGAATTCGG-3’; IFN-β-F, 5'- AAGAGTTACACTGCCTTTGCCATC-3’, IFN-β-R, 5'- CACTGTCTGCTGGTGGAGTTCATC-3’; IL-6-F, 5'-GACAAAGCCAGAGTCCTTCAG AGAG-3’, IL-6-R, 5'-CTAGGTTTGCCGAGTAGATCTC-3’; IFN-α-F, 5'-CGCAGGAGAAGGTGGATGCCCAG-3’, IFN-α-R, 5'-CAGCACATTGGCAGAGGAAGACAGG-3’ [19 (link)].
The mRNA level of TNF-α, IFN-β, IL-6, and IFN-α in GM-DCs were tested with SYBR green method as described above. Primers used for the cytokine test were the following:
TNF-α-F, 5'-GGCAGGTCTACTTTGGAGTCATTG-3’, TNF-α-R, 5'-ACATTCGAGGCTCCAGTGAATTCGG-3’; IFN-β-F, 5'- AAGAGTTACACTGCCTTTGCCATC-3’, IFN-β-R, 5'- CACTGTCTGCTGGTGGAGTTCATC-3’; IL-6-F, 5'-GACAAAGCCAGAGTCCTTCAG AGAG-3’, IL-6-R, 5'-CTAGGTTTGCCGAGTAGATCTC-3’; IFN-α-F, 5'-CGCAGGAGAAGGTGGATGCCCAG-3’, IFN-α-R, 5'-CAGCACATTGGCAGAGGAAGACAGG-3’ [19 (link)].
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