Either miR-498 or miR-320d regulates KSHV replication in BCBL-1 cells without HSV-1 infection.

<p>(<b>A</b>)<b>. Transfection of miR-498 or miR-320d mimic inhibited KSHV RTA expression in TPA-stimulated BCBL-1 cells.</b> Western blot was used to detect the expression of RTA in BCBL-1 cells which were transfected with miR-498, miR-320d mimics or Neg. Ctrl. for 12 h, and stimulated with TPA for another 24 h. The relative level of RTA was determined by quantitative densitometry. Numbers labeled above the RTA band were the relative intensities of the bands compared to GAPDH. The relative level of RTA in the TPA+Neg. Ctrl. group was considered to be 1 for comparison. (<b>B</b>)<b>. Transfection of miR-498 or miR-320d mimic inhibited KSHV ORF21/57 mRNA expression in TPA-stimulated BCBL-1 cells.</b> ORF21 and ORF57 mRNA in BCBL-1 cells treated as in A was quantitated by RT-qPCR. *<i>P</i><0.05, **<i>P</i><0.01 and ***<i>P</i><0.001 for Student’s t-test versus TPA+Neg. Ctrl. group. (<b>C</b>)<b>. Transfection of miR-498 or miR-320d mimic inhibited KSHV progeny virions release in TPA-stimulated or unstimulated BCBL-1 cells.</b> Real-time DNA-PCR was used to detect the viral genome copy number in the supernatant of BCBL-1 cells, which were transfected with miR-498, miR-320d mimics or Neg. Ctrl. for 12 h, and treated with TPA for another 24 h (left columns) or without TPA (right columns). *<i>P</i><0.05, **<i>P</i><0.01 and ***<i>P</i><0.001 for Student’s t-test versus Neg. Ctrl. group. (<b>D</b>)<b>. Transfection of LNA-498 or LNA-320d enhanced KSHV RTA expression in TPA-stimulated BCBL-1 cells.</b> Western blot was used to detect the expression of RTA in BCBL-1 cells transfected with LNA-498, LNA-320d or LNA-Scr. Ctrl. for 12 h, and stimulated with TPA for another 24 h. The relative level of RTA was determined by quantitative densitometry. Numbers labeled above the RTA band were the relative intensities of the bands compared to GAPDH. The relative level of RTA in the TPA+LNA-Scr. Ctrl. group was considered to be 1 for comparison. (<b>E</b>)<b>. Transfection of LNA-498 or LNA-320d enhanced KSHV ORF21/57 mRNA expression in TPA-stimulated BCBL-1 cells.</b> ORF21 and ORF57 mRNA in BCBL-1 cells treated as in D was quantitated by RT-qPCR. *<i>P</i><0.05, **<i>P</i><0.01 and ***<i>P</i><0.001 for Student’s t-test versus TPA+LNA-Scr. Ctrl. group<b>.</b> (<b>F</b>)<b>. Transfection of LNA-498 or LNA-320d enhanced KSHV progeny virions release in TPA-stimulated BCBL-1 cells.</b> Real-time DNA-PCR was used to detect the viral genome copy number in the supernatant of BCBL-1 cells treated as in D. ***<i>P</i><0.001 for Student’s t-test versus TPA+LNA-Scr. Ctrl. group.