Pathogens used to evaluate specificity.

<p>Pathogens used to evaluate specificity.</p

Real-time RT-PCR mix composition.

<p>*In the case of two primers/probes, this is a 50:50 mix of each primer/probe</p><p>Real-time RT-PCR mix composition.</p

Specificity of pan-EHDV and serotyping assays.

<p>Specificity of pan-EHDV and serotyping assays.</p

Real-time RT-PCR validation.

<p>* CI: confidence interval CV: coefficient of variation ND: Not done.</p><p>Real-time RT-PCR validation.</p

Transcriptional response to CAV2 vector in the CD103⁺ and CD11b⁺ -type DC subsets.

<p>(A)The mean fold induction of the selected genes up-regulated by Cav-null R⁰ are represented in graded yellow to red color (from 1 to >3) for both CD103⁺ and CD11b⁺ -type DCs from 4 sheep. The represented genes were selected from the significantly induced genes in the CD11b⁺ -type DC subset (222 genes, p<0.05, fold > 2) and they were ranked based on their fold induction in the CD11b⁺ -type DC subset. The significantly activated genes in the CD103⁺ subset (21 genes, p<0.05, fold > 2) are indicated by a star. (B) The mean fold reduction levels of the selected down-modulated genes by Cav-null R⁰ are represented in graded yellow to blue color (from 1 to <0.2) for both CD103⁺ and CD11b⁺ -type DCs from 4 sheep. The represented genes were selected from the significantly reduced genes in the CD11b⁺ -type DC subset (29 genes, p<0.05) and they were ranked based on their fold induction in the CD11b⁺ -type DC subset. No gene expression was significantly reduced in the CD103⁺ -type DC subset.</p

Antimicrobial gene network induced by CAV2 vector in CD11b⁺ -type DCs.

<p>An antimicrobial gene network centered on IFN was generated by the Ingenuity Pathways Analysis on the selected genes dys-regulated by CAV2 vector in CD11b⁺ -type DCs (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0052513#pone.0052513.s003" target="_blank">Table S1</a>, p < 0.05 and fold > 2, or p < 0.05 and folds confirmed by qRT-PCR). Molecule types are represented by symbols: diamonds (enzymes), triangles (kinase), square (cytokine), double circle (complex), oval (transcription regulator), circle (others).</p

Top canonical pathways induced by CAV2 vector in CD103⁺ and CD11b⁺ -type DCs.

<p>Top canonical pathways induced by CAV2 vector in CD103⁺ and CD11b⁺ -type DCs.</p

Validation of gene expression induced by CAV2 vector with qRT-PCR.

<p>A selection of genes was checked for CAV2-induced upregulation using qRT-PCR on cDNA from the CD103⁺ and CD11b⁺ -type DCs (4 different sheep were used in total, 3 per validated gene). Each gene detection was normalized with GAPDH expression and the relative gene expressions (log2(CTgene-CTGAPDH)) are reported. The gene names printed with regular font were selected as being up-regulated in the CD11b⁺ type DC microarrays (p < 0.05 ANOVA followed by Benjamini-Hochberg FDR and > 2 fold), the gene names printed in italic are not represented by any probe on the ovine microarray, the underlined gene names were close to be selected by the variance analysis in the CD11b⁺ -type DCs.</p

Biological functions that are predicted to be activated by CAV2 vector in CD103⁺ and CD11b⁺ -type DCs.

*<p>the expression of the underlined molecules is down-modulated.</p

Expression of VP7 antigen.

<p>Following transduction of CHO-CAR cells with Cav-VP7 R⁰, expression of bluetongue antigen was sought and detected by immunostaining using a MAb directed against the VP7 protein (A). Following transduction of RK13 cells with SG33-VP7, VP7 antigen was labeled using an anti-VP7 hyperimmune rabbit serum (C). Non-transduced cells were used as negative control (B and D).</p