Figure 3. The effect of S. purpurea extracts on VACV transcription in vivo and in vitro.

A) HeLa cells were infected with VACV at an MOI = 10 followed by the addition of 25 microL S. purpurea extract/ml media. At 4 HPI, total RNA was isolated and VACV-E3L RNA levels determined by real-time PCR. Reactions contained total RNA from mock-infected cells, ethanol/glycerol carrier-treated VACV-infected cells, or S. purpurea-treated VACV-infected cells. C(t) values were calculated using manufacturer's software. Graph illustrates data from a representative experiment. C(t) values and fold change from two separate experiments are shown. B) Purified VACV virion cores were incubated in the presence of the indicated concentrations of S. purpurea or ethanol/glycerol carrier and [³⁵S]-UTP. Newly synthesized RNA products were spotted onto glass-fiber filters, washed in TCA, and quantified by scintillation counting (counts per minute). A no template reaction was performed by excluding the addition of the virion cores.