Fig 4.

(A) Stability of BG-323 at 37°C. DMSO or 50 μM (final concentration) BG-323 was added to the culture medium (DMEM), and medium samples were incubated in a CO₂ incubator at 37°C for 48 or 24 h prior to infection. At time zero, preincubated medium or fresh medium and BG-323 were added to BHK cells and the cells were subsequently infected with Kunjin virus at an MOI of 0.01. At 72 h postinfection, medium samples were collected and stored at −80°C. RNA was prepared and quantified by qRT-PCR as described in Materials and Methods. n = 3. (B) Replacement of BG-323 reduces Kunjin virus replication. BHK cells were infected with Kunjin virus at an MOI of 0.01 and treated with 100 μM BG-323 or DMSO. At 24, 48, and 72 h postinfection, medium samples were collected and medium on infected cells was replaced with fresh medium with 100 μM BG-323 or DMSO as indicated. Viral RNA at 24, 48, and 72 h postinfection (PI) was quantified by qRT-PCR as PFU equivalents/ml. n = 3.