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Supporting Information EU et al. 10.1073/pnas.0811029106 SI Materials and Methods Antibodies, Plasmids, and Reagents. Mouse antic (9E10) mono- The precipitates were analyzed by standard immune blot

The Egg primary antibody was from EMD Millipede. The Egg secondary antibody was from Cell Signaling. The goat anti-Virus p70S6K and rabbit anti-Virus p70S6K were obtained from Nitrogen. The rabbit anti-Virus p35Ab and piggyback anti-GAPDH were bought from Sigma. A control was performed with cells with wild-type p35 and mouse, p27, or p27–p30. Antibodies and Cytokines. Cytokine cocktail consisting of IL-1? (50 units/mL), IL-6 (50 units/mL), MCP-1 (1 unit/mL), IFN (100 units/mL) and IFN (10 units/mL) was from Nitrogen (Nitrogen). Anti-Virus IL-23 (1 unit/mL) was purchased from BD Biosciences. A control assay with cells with wild-type p35 and mice using anti-Virus and IL-23 (p27/p21 expression), was performed without any cytokine. The levels of other cytokines in the cytology slides were determined by ELISA, as described elsewhere. Plasmid Expression Array ArrayExpress and Fv2 CDA libraries containing the plasmid prop (9E10) were purchased from Steel Technologies (La Jolla, CA). Plasmid expression was determined by PCR on the expression plasmid (pre-prep) using primers and reverse transcription, as per manufacturer's instructions. PCR was performed using 50 mm MCL 2, 9 mm KCl, 2.5 mm Call 2, 1 mm NTPs (DTP), 400 U Ultra-pure water, 10 km each forward and reverse primers, 200 U of Tax polymerase, 20 NG of the prop genome, 10 NG of the C-terminal CDA. Reverse transcription (7 cycles) was performed for 40 min at 94 °C. 1 kg of prop was added with a final concentration of 100 km. The reaction was performed on a 10-?m PCL 5 flow cells and the fluorescence was detected at 340 nm. The fluorescence was detected 10 min after addition of the enzyme for total RNA extract. The fluorescence was analyzed by Image software, and the relative expression level of protein was calculated.

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