FIG 3.

RTA is required for KDM2B-induced lytic gene expression and viral DNA replication during de novo infection. (A and B) SLK cells were transduced with lenti-GFP or lenti-KDM2B for 3 days, followed by KSHV infection for 3 days in the presence or absence of 100 μM PAA. (A) Viral DNA load was measured by qPCR at 72 hpi relative to 2 hpi. (B) RT-qPCR analysis of viral gene expression at 72 hpi. (C to E) SLK cells transduced with lenti-GFP or lenti-3×FLAG-KDM2B for 3 days followed by infection with WT or RTA-KO KSHV for 3 days. (C) Viral DNA level at 72 hpi relative to 2 hpi was measured by qPCR. (D) RT-qPCR detection of viral gene expression in KDM2B-OE cells relative to GFP-OE cells at 72 hpi. (E) Immunoblot detection of lytic viral protein production in WT and RTA-KO KSHV-infected GFP-OE and KDM2B-OE cells at 72 hpi. Tubulin was used as a loading control. t test was performed between (−PAA) and (+PAA) samples (A and B) as well as between GFP-OE and KDM2B-OE samples (D), and P < 0.05 (*) was considered statistically significant. Asterisks at the immunoblots indicate nonspecific signal.