Persistent prion infection disturbs the function of Oct-1, resulting in the down-regulation of murine interferon regulatory factor-3

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

Next, we challenged with 0.1% brain homogenates from 22Linfected mice (22LBH) or normal mice (nBH) to N2a58 cells and incubated for 48 h. As expected, the promoter activity in 22LBHincubated cells was significantly decreased ( Supplementary Fig. S1 ), indicating that prion infection reduced the activity. Furthermore, drug treatment was started after the initial seeding of the cells (at 75 cm 2 flask) and renewed every passage. The significantly reduced promoter activity in ScN2a58 cells increased after continuous treatment with Congo red (CR, 10 mM) or pentosan polysulfate (PPS, 20 mg/ml), which are both known as anti-prion drugs 12 (Fig. 2c ; CR: 3.8-fold and PPS: 3.4-fold). The efficacy of the two drugs was evaluated by detection of PK-resistant PrP using western blotting in cells after final passage (Fig. 2d) . The activities of the uninfected control N2a58 cells were not affected during continuous treatment ( Supplementary Fig. S2 ).