Type I interferon protects neurons from prions in in vivo models

Ishibashi, Daisuke; Homma, Takujiro; Nakagaki, Takehiro; Fuse, Takayuki; Sano, Kazunori; Satoh, Katsuya; Mori, Tsuyoshi; Atarashi, Ryuichiro; Nishida, Noriyuki

Polyinosinic-polycytidylic acid (Poly I:C, Invivogen), Pentosan polysulfate (PPS, Sigma-Aldrich) and RO4948191, which is an imidazonaphthyridine with the structural formula 8-(1, 3, 4-oxadiazol-2-yl)-2, 4-bis (trifluoromethyl) imidazo [1, 2-a] [1, 8] naphthyridine (BIONET: RO8191), were purchased from the indicated vendors. Recombinant mouse IFN-and -b were purchased from Calbiochem (Merck). The mouse monoclonal antibody 3S9, recognizing the helix 1 region of PrP, was a kind gift of Prof. Matsuda, Hiroshima University (Miyamoto et al., 2005) . The anti-PrP polyclonal mouse antiserum (SS), M20 (Santa Cruz Biotechnology), and the monoclonal (SAF32 and SAF83, SPI-Bio) antibodies have been described previously (Atarashi et al., 2006; Ishibashi et al., 2007 Ishibashi et al., , 2011 Homma et al., 2014b) . Anti-IFNAR1 (MAR1-5A3, BioLegend), anti-IFN-b (7F-D3, Santa Cruz Biotechnology), anti-ionized calcium binding adaptor molecule 1 (Iba-1; Wako Pure Chemical Industries, Ltd), anti-glial fibrillary acidic protein (GFAP; Dako), anti-murine 2'-5' oligoadenylate synthetase (OAS1a; C-5, Santa Cruz Biotechnology), anti-HA (Invitrogen), and b-actin (Sigma) were mouse, rat, and rabbit monoclonal antibodies, and were purchased from commercial sources. The horseradish peroxidase-conjugated anti-goat (Jackson ImmunoResearch Laboratories), antirat (Cell Signaling Technology, Inc.: CST), anti-mouse, and antirabbit (GE healthcare Life Sciences) IgG antibodies were used for immunoblotting.