Expression of KSHV lytic cycle RNA and protein in HSV-2-infected BCBL-1 cells.

<p>(<b>A</b>). <b>Rta mRNA expression in HSV-2-infected BCBL-1 cells.</b> Rta mRNA expression in BCBL-1 cells infected with Mock or HSV-2 for 3, 6, 12, 24, 48, 72 and 96 hours was quantitated by RT-qPCR. Relative quantities of Rta expression represented on the y-axis. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031652#s2" target="_blank">Results</a> shown were from three independent experiments performed in triplicate. * <i>p</i><0.05 and ** <i>p</i><0.01 for Student's t-test versus mock group, respectively. (<b>B</b>). <b>ORF26 mRNA expression in HSV-2-infected BCBL-1 cells.</b> ORF26 mRNA expression in BCBL-1 cells infected with Mock or HSV-2 for 3, 6, 12, 24, 48, 72 and 96 hours was measured by RT-qPCR. Relative quantities of ORF26 expression represented on the y-axis. Results shown were from three independent experiments performed in triplicate. * <i>p</i><0.05 and ** <i>p</i><0.01 for Student's t-test versus mock group, respectively. (<b>C</b>). <b>RT-PCR analysis for ORF57 and vIL-6 mRNA in BCBL-1 cells infected with HSV-2.</b> ORF57 and vIL-6 mRNA expression in BCBL-1 cells infected with Mock or HSV-2 for 24, 48, 72 and 96 hours was detected by RT-PCR. β-Actin was readily detectable in all samples indicating the presence of amplifiable cDNA. (<b>D</b>). <b>Rta protein expression in BCBL-1 cells infected by HSV-2.</b> BCBL-1 cells were infected with HSV-2 or mock for 24, 72 and 96 h and whole-cell lysates were subjected to Western blot with Rta antibody. The membrane was stripped and reprobed with anti-actin antibody as a loading control. Results shown are from a representative experiment of three independent experiments with similar results. (<b>E</b>). <b>Immunofluorescence assay staining of BCBL-1 cells infected with HSV-2 (original magnification, 40×).</b> KSHV lytic protein ORF59 expression in normal BCBL-1 cells (untreated) and BCBL-1 cells infected with mock or HSV-2 for 24 and 72 h was detected by immunofluorescence assay staining with ORF59 MAb. (<b>F</b>). <b>Quantification of results in</b> (<b>E</b>). Quantitative and statistic methods were described in the text. ** <i>p</i><0.01 for Student's t-test versus mock group.