Recombinant human ll 37

MitoTracker Green, MitoTracker DeepRed, MitoSox, and CellRox were purchased from Molecular Probes. Proteinase K was purchased from Roche. Recombinant human GM-CSF was purchased from BD. Recombinant human LL-37, R837, ODN-TTAGGG, and ODN-2216 were purchased from InvivoGen. FcR Blocking Reagent was purchased from Miltenyi Biotec. Antibody against 8OHdG (J-1; rabbit polyclonal IgG2b) and all chemicals were purchased from Santa Cruz Biotechnology, Inc. Antibodies against LL-37, TFAM (Clone 18G102B2E11), Mitofilin (Clone 2E4AD5), γH2A.X, TOMM20 (Clone 4F3), Pyruvate Dehydrogenase E2/E3bp (PDH), VDAC/Porin, Glyceraldehyde 3-phosphate dehydrogenase (GADPH), dsDNA, LAMP1 (Clone H4A3), and Rab7 were purchased from Abcam. Antibodies against MnSOD were purchased from Millipore. Antibodies against HMGB1, PGC1α, and Phospho-(Ser/Thr) PKA Substrate were purchased from Cell Signaling Technologies. Genomic DNA was obtained from BioChain. IRS661 and DVX41 were a gift from Dynavax Technologies Corporation. Specified neutrophils were cultured in the presence of αRNP-IgG (50 µg/ml), Recombinant human GM-CSF (50 ng/ml), diphenylene iodonium (DPI; 10 µM), MT (10 µM), CCCP (25 µM), Rotenone (5 µM), Oligomycin (10 µM) plus Antimycin (1 µM; O+A), IRS661 (TLR7 antagonist; 1 µM), DVX42 (TLR8 antagonist; 1 µM), Bafilomycin A1 (BafA1; 1 µM), 8Br-cAMP (100 µM), IBMX (50 µM), or H89 (1 µM).