Figure 4. Effect of HM on IE gene expression.

[A] Western blot analysis: 40 µg of protein sample from whole cell extracts were separated by SDS-PAGE, blotted to nitrocellulose, and filters were incubated with monoclonal anti-ICP4 or polyclonal anti-β actin antibody followed by decoration with peroxidase-labelled anti-rabbit polyclonal antibodies, respectively and visualized using ECL Western blot detection kit. [B] Quantitative real time PCR: HSV-2G (5 moi) infected cells were treated with HM (5.0 µg/ml) for 2 and 4 h. After incubation at 37°C RNA was isolated and subjected to cDNA synthesis. Then quantitative real-time PCR was performed with these products by using SYBR Green PCR Master Mix. PCRs were amplified with the cycling conditions of: 95°C for 10 min and 40 cycles (15 s at 95°C, then 60 s at 60°C).