Figure 9.

GPS491 inhibits 229E virus replication post entry. (a) Kinetics of 229E coronavirus N protein expression and virus release. Huh7 cells were infected at an input MOI of 2. After incubation of cells for 1 h, cells were washed to remove residual virus and fresh media were added. At indicated times post infection (p.i.), cells and media were harvested. Cell extracts were analyzed by Western blot for N protein expression, while RTqPCR was used to monitor levels of viral RNA in the media. Shown is a representative Western blot as well as a graphical summary of the quantitative results obtained from n = 3 independent assays. (b) Addition of GPS491 at late stages of 229E replication suppresses virus growth. Huh7 cells were infected at an input MOI of 2. After incubation of cells for 1 h, cells were washed to remove residual virus and fresh media were added. At indicated times post infection (p.i.), DMSO or GPS491 was added to cultures at a final concentration of 1.25 µM and incubation continued. At 24 h post-virus infection, media were harvested and the abundance of viral RNA in media was determined by RTqPCR.