Figure 5.

Effect of modulated ERK pathway on HSV-1 replication. (a) MOI-dependent growth curves were obtained by infecting HEp-2 and HEp-dnERK with HSV-1 at MOI 1 and 10. The sample were harvested at 1 h, 18 h and 36 h pi, the virus yields were measured by standard plaque assay on Vero cells. The procedure was described in Methods. (b) HEp-2, HEp-dnERK and HEp-dnERK 4/2 cell lines were infected with HSV-1 at MOI 10 and collected 24 h pi. The viral DNA was extracted 24 h post HSV-1 infection as described in the Methods. Viral DNA amounts were determined by absolute real-time PCR using a TaqMan probe. The result is expressed as concentration in ng of DNA/µl. (c–f) Quantitative Real-Time PCR of gB, ICP0, CyclinE and CDK2 transcripts in HEp-2 and HEp-dnERK cells were evaluated at 3 h, 9 h, 24 h pi. Calculations of the copies number were based on external standards and all transcripts were normalized against the GAPDH copies number. Statistical analyses were performed with one-way ANOVA analysis assay in triplicate and *P ≤ 0.05, **P < 0.01 ***P < 0.001 indicate significant change”.