Figure 3. The canonical nuclear import α/β 1 pathway is important for infection.

(A) RPTE cells were infected at an MOI of 10⁴ genomes/cell and treated with ivermectin (+I) or butanol (untreated, UT). Whole cell lysates were harvested at 24 hpi and resolved by SDS-PAGE, then probed for TAg and β actin. (B) RPTE cells were infected with 10⁴ genomes/cell of wild-type (Dun) or mutant (DunM) and treated with ivermectin (+I) at 10 μM at the beginning of infection or left untreated. RNA was harvested at 24 hpi and reverse-transcription (RT) was performed followed by qPCR to measure TAg cDNA relative to GAPDH. (C) RPTE cells were transfected with Importin β1 siRNA (Imp β) or non-targeting siRNA (N.T.), and then infected 2 d after transfection with wild type virus. Whole cell lysates were harvested at 24 hpi, resolved by SDS-PAGE, and probed for TAg, B actin, and importin β1. (D) Transfection and infection was performed as in C, and RNA was harvested at 24 hpi. RT and qPCR was performed as before. Results represent three independent experiments.