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Upon infection with PI-3 virus, cultures showed a decrease in cell growth and MC loading directly related to the multiplicity of infection (moi) used for virus infection.
T3 homozygous lines were used for virus inoculation.
A five-tube mRT-PCR was used for virus identification as previously described [43], [44].
The remaining swab lysate was used for virus isolation (see below).
Lentiviral vectors were constructed and used for virus generation as described previously [5], [6].
BHK-21 and Vero cells used for virus production and titration were cultured under similar conditions.
Ultracentrifuge-purified BYDV-byd1 virus (fifth passage on DEF monolayer) was used for virus genome amplification.
One ml of the washed RBC was also used for virus isolation in C. sonorensis (KC) cells.
About 100 ml of the homogenized sample was used to extract genomic DNA (AquaPure DNA isolation kit) and the remaining volume was used for virus isolation (see below).
To prepare cells for these assays, 80% confluent cells were treated with RBV for 24 h (same conditions used for virus infections in Figure 1).
Triton x-100 was used for virus inactivation.
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