Generation, characterization and integration efficiency of the ΔimpTMR mutant.

<p><b>(A)</b> Schematic representation of wt, ΔimpTMR and ΔimpTMRrev. <b>(B)</b> Southern blot analysis detecting TMR sequences of the impTMR (lower panel) and pTMR (upper panel) after <i>Eco</i>RV digestion of the indicated BAC clones using a DIG-labeled TMR probe. <b>(C)</b> Growth kinetics comparing replication properties of wt, ΔimpTMR mutant and ΔimpTMRrev virus in JJHan cells. HHV-6A genome copy numbers were detected by qPCR. Copy numbers per 1x 10⁶ cells are shown as means of three independent experiments with corresponding standard errors. <b>(D)</b> Integration frequency was quantified by determining the integration status of at least 90 metaphases. Significant differences between wt and ΔimpTMR, as well as wt and ΔimpTMRrev (Mann-Whitney U-test, p < 0.05) are indicated with an asterisk (*). Results are shown as means of three independent experiments with standard errors. <b>(E)</b> 300 interphase nuclei were examined for the presence of HHV-6A. Significant differences between wt and ΔimpTMR, as well as wt and ΔimpTMRrev (Mann-Whitney U-test, p < 0.05) are indicated with an asterisk (*). Results are shown as means of three independent experiments with standard errors. <b>(F)</b> Maintenance of the HHV-6A genome was determined by qPCR analysis at d0 and d14 post sorting. Copy numbers per 1x 10⁶ cells are shown as means of three independent experiments with standard errors. Significant differences between wt and ΔimpTMR, as well as wt and ΔimpTMRrev (Mann-Whitney U-test, p < 0.05 or p < 0.01) are indicated with asterisks (* or **).