FIG 4.

E8 mutations increase viral early transcription in differentiated organotypic cultures. (A) Total RNA from 12-day HPV16 wt (wt), HPV16 E8− (E8−), and HPV16 E8 KWK mt (E8 KWK mt) organotypic cultures was analyzed by qRT-PCR for the expression of E6, E7, E1, and E2 transcripts using PGK1 as a reference gene. Copy numbers are derived from plasmid dilutions of pBS 16 114b:1 and hPGK1 pEF-IRES-P plasmids measured in parallel. The data represent averages from six independent organotypic cultures, and error bars show the standard errors of the means. Statistical significance between results for the wt and mutants was determined by paired, two-sided Student's t test (∗, P < 0.05; ∗∗, P < 0.01). The differences between E8− and E8 KWK mt are not significant. (B) Relative changes of viral transcript levels (E6, E7, E1, and E2) induced by differentiation in HPV16 wt, E8−, or E8 KWK mt genome cell lines.