Elongation of O-linked glycans affects HSV-1 secretion/infectivity.

<p>(A) HaCaT wild-type or HaCaT mutant keratinocytes, lacking elongation of mucin-type O-liked glycosylation (HaCaT sc D5 and HaCaT sc E5) were infected with MOI of 10 of HSV-1 Syn17+ produced in HaCaT wt. Medium was harvested at 12 and 20 hours post-infection and number of infectious particles were quantified using plaque titration on Vero culture monolayer and expressed as plaque forming units per mL of medium (PFU/mL). Bar graphs represent mean values of 3 biological replicates assayed by 2 technical replicates each + SD. A 2-way ANOVA with Tukey’s multiple comparison test was used to compare differences between means. NS—<i>p</i> > 0.05, *—<i>p</i> < 0.05, **—<i>p</i> < 0.01, ***—<i>p</i> < 0.001, ****—<i>p</i> < 0.0001. Results are representative of at least two independent experiments. (B) Numbers of viral DNA copies in the medium were quantified by qPCR and using a standard curve based on amplification of known copy numbers of HSV-1 DNA fragments cloned in Topo TA plasmids. Copy numbers of viral DNA are expressed as copies/mL. Bar graphs represent mean values of 3 biological replicates assayed by 3 technical replicates each + SD. A 2-way ANOVA with Tukey’s multiple comparison test was used to compare differences between means. NS—<i>p</i> > 0.05, *—<i>p</i> < 0.05, **—<i>p</i> < 0.01, ***—<i>p</i> < 0.001, ****—<i>p</i> < 0.0001. Results are representative of at least two independent experiments.