Fig. 3.

STIV infection in S. solfataricus PH1-16 expressing the trans-dominant negative Vps4 Walker B mutant. (A) qPCR measurements of STIV B345 gene abundance during the course of STIV infections and mock infections in S. solfataricus PH1-16 cells repressing or expressing a Walker B mutant form of Vps4. (B) Western blot analysis of episomal Vps4 Walker B mutant expression. Episomal Vps4 was detected with the anti-FLAG antibody (Upper) and the TATA Binding Protein (TBP) served as a loading control (Lower). Cultures were induced (I) with arabinose or repressed (R) with galactose 24 h after STIV infection. (C) RT-PCR of RNA extracted from S. solfataricus PH1-16 cultures infected with STIV shows transcription of an early viral gene product (C121). Samples were collected at 24 hpi, and the extracted RNA was subjected to a one-step RT-PCR. The Taq-only control was performed without reverse transcription to test for contaminating DNA. NTC, no template control; size standard is in base pairs.