Genome-wide analysis of regulatory G-quadruplexes affecting gene expression in human cytomegalovirus
Fig 7
Effects of G4-binding ligands on viral gene expression, DNA replication, and production of progeny virions.
(A) Relative mRNA levels in response to G4-binding ligands. HF cells were infected with HCMV (Toledo) at an MOI of 1, and the mRNA levels were measured by qRT-PCR at 24 h (for immediate-early genes), 32 h (for early genes) or 48 h (for late genes). To examine the effect of G4-binding ligands, cells were treated with 5 μM of NMM and TMPyP2 for 24 h prior to cell harvest. The relative mRNA levels normalized by those of β-actin are shown. (B and C) Comparison of viral DNA levels and viral titers in response to G4 ligands. HF cells were infected with HCMV (Toledo) at an MOI of 1 for 72 h. Cells and supernatants were separately collected. The viral DNA levels from the cells (intracellular) and the supernatants (extracellular) were determined by qPCR. The relative amounts of viral DNA levels normalized by those of β-actin are shown (B). The cell-associated and cell-free virus titers were also determined using infectious center assays (C).