Various G-CSF concentrations with conditioned medium were assayed by using Ready-Set-Go ELISA kit (R&D systems) as described by the manufacturer.
The synthesis of first strand cDNA was carried out from 1 µg of total RNA by using Ready-To-GoTM RT-PCR beads (GE Healthcare, UK) and the manufacturer's protocol.
Random amplified polymorphic DNA (RAPD) analysis was performed by using Ready-To-Go RAPD analysis beads (Amersham Pharmacia Biotech, Piscataway, New Jersey), according to the manufacturer's instructions.
We then extracted the total RNA from 140 µL of supernatant by using the QIAamp Viral RNA Mini Kit (QIAGEN, Valencia, CA, USA) in accordance with the manufacturer's protocol and produced the first strands of cDNA by using Ready-To-Go You-Prime First-Strand Beads (Amersham Pharmacia Biotech, Piscatawy, NJ, USA) as described in the manual accompanying the kit.
RT was performed by using Ready-To-Go-You Prime First Strand Beads (Amersham Pharmacia Biotech, Piscatawy, NJ, USA) and a seminested PCR to amplify 492-bp gene fragments of the premembrane (PrM) sequence of JEV by using the Takara LA Taq PCR kit (Takara Bio Inc., Shiga, Japan).
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cDNA was prepared by using Ready-to-Go You-Prime First-Strand Biotechit (Amersham Piscatawy Biotech, Piscatawy, NJ, USA) according to the manufacturer's protocol.
The activity of specific proteases was assessed by zymography using Ready Gel Zymogram Gels containing gelatin or casein (BioRad Laboratories, Hercules, CA).
First strand cDNA was generated from total RNA by RT using Ready-To-Go You-Prime first strand beads (GE Healthcare, Buckinghamshire, UK) following manufacturer's instructions.
IFNγ plasma levels were determined by ELISA using Ready-Set-Go kits with precoated plates (eBioscience, San Diego, CA, USA) as per the manufacturer's protocol.
cDNA containing the CFP tag sequence was amplified by nested PCR using Ready-Mix PCR mix (Bio-Lab).
