FIG 6.

p53 promotes prereplication ICP8, but not ICP4 or ICP27, gene transcription. Cas9 and TP53 #5 cells were infected with 7134 at an MOI of 1, and nucleic acids were collected 24 h later. Purified DNA was amplified and quantified via real-time PCR with primer pairs for the UL29 locus of the viral genome and cellular GAPDH. The values for UL29 were normalized to the GAPDH values (A). cDNAs of viral genes representative of each viral gene class were quantified. ICP4 and ICP27 represented the IE class, ICP8 the E class, and gC the L class. Cells were treated with ACV to allow the quantification of IE and E transcripts before the onset of viral DNA replication (B). Total RNA from the same samples collected for panel A was isolated and reverse transcribed, and the indicated viral transcripts were quantified via real-time PCR and normalized to cellular 18S rRNA transcripts (C). Data are averages with SEM from three independent experiments (A and C). Statistical significance was determined, compared to the Cas9 control cells, via unpaired t tests. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.