Figure 2.

JΔNI5ΔB and JΔNI8ΔB Transgene Expression in HDFs and rDRGs

(A) mCherry transgene expression in rDRG cultures. Cells were infected with 3,000 gc/cell of JΔNI5ΔB (top) or JΔNI8ΔB (bottom). Bright-field (left) and mCherry fluorescence images (right) were taken at 7 (left panels) and 14 dpi (right panels). (B) mCherry mRNA levels in JΔNI5ΔB- and JΔNI8ΔB-infected rDRGs at 7 and 14 dpi. mCherry mRNA levels were normalized to viral gc in the same samples, and the normalized values are presented relative to JΔNI5ΔB-infected cells at 7 dpi. (C) Cytotoxicity for rDRGs in culture. Cells were infected with KOS, JΔNI5ΔB, or JΔNI8ΔB virus at 3,000 gc/cell or mock-infected, and cell viability was measured in triplicate by MTT assay at 5 dpi. Plotted values represent the mean ratios of virus-infected to mock-infected cells. (D) Transgene and viral gene expression in HDFs (left) and rDRGs (right). HDF and rDRG cultures were infected with JΔNI5ΔB or JΔNI8ΔB vectors at 25,000 gc/cell or 3,000 gc/cell, respectively. At 7 dpi, mRNA levels of selected genes were measured by qRT-PCR analysis. Data conversion was performed using JΔNI5 BAC DNA standard curves generated with the same gene-specific primers and normalization to viral gc in each sample. Values are presented relative to JΔNI5ΔB-infected HDF at 7 dpi. Data in (B) and (D) represent averages ±SD of two independent experiments.