Figure 1.
JΔNI5 Vector Growth at Different MOIs
Confluent U2OS-ICP4/27 cells in T225 flasks were infected with JΔNI5 vector at 10−2–10−6 plaque formation unit (PFU)/cell. (A and B) Physical titers in gc/mL (A) and biological titers in PFU/mL (B) in the supernatants were measured daily by quantitative real-time PCR and standard plaque assay, respectively. (C) gc/PFU ratios were calculated based on the results of (A) and (B). (D and E) Cultures of rat primary dorsal root ganglia (rDRGs) were infected with JΔNI5 vector preparations (5,000 gc/cell) produced at MOIs of 10−2 or 10−5. At 4 d post-infection (dpi), vector-mediated mCherry fluorescence was photographed (D) and relative mCherry mRNA levels were measured by qRT-PCR with normalization to 18S rRNA (E). Group comparisons were performed with Student’s t test or one-way ANOVA with post hoc Dunnett’s multiple comparison test. Differences between an input MOI of 10−5 and 10−2 were statistically highly significant (∗p < 0.05). Quantitative data are presented as means ± SD (n = 3).