Fig 6.

Effects of KDM3A knockdown on the KSHV life cycle. (A) Viral gene expression. KSHV reactivation was triggered by incubation with doxycycline for 4 h. Total RNA was harvested at the indicated time points, and KSHV PCR array analysis (a) or individual RT-qPCR (b) was performed to monitor viral gene expression. Cellular GAPDH and actin were used for internal controls, and the mean normalized expression (MNE) level is shown. For array analyses, individual gene expression levels were normalized to the lowest expression level for each column, and fold induction of viral gene expression is shown as a heat map. (B) Viral protein expression. A series of immunoblot analyses was performed to examine the expression of KSHV immediate early, early, and late genes at the protein level. The indicated antibodies were used to probe viral protein expression. Cellular GAPDH was used as a loading control. (C) Viral replication. The intracellular viral DNA copy number was measured by qPCR. The actin coding region was used as an internal control.