Relative Log2 transformed gene expression levels of the three subsets and statistical significance the subsets in between.

<p>C = Classical, I = Intermediate, NC = Non-Classical</p><p>Relative Log2 transformed gene expression levels of the three subsets and statistical significance the subsets in between.</p

Genes of interest investigated in the three monocyte subsets.

<p>Protein function adapted from <a href="http://GeneCards.org" target="_blank">GeneCards.org</a></p><p>Genes of interest investigated in the three monocyte subsets.</p

Expression level of genes deviating in identified subgroups.

<p>Subgroups of cells were identified based on the PCA of single-cell PCR gene expression analysis data. One subgroup among the classical monocytes, intermediate monocytes and non-classical monocytes, and co-expression of genes within the subgroups were assessed using a student T test (<i>P <</i> 0.05). A) Bar graph demonstrating the differentially expressed genes by the subgroup within the classical monocyte subset identified on the PCA score plot. The subgroup is marked by filled red circles in the PCA score plot. B) Bar graph demonstrating the differentially expressed genes by the subgroup within the intermediate monocyte subset identified on the PCA score plot. The subgroup is marked by filled green triangles in the PCA score plot. C) Bar graph demonstrating the differentially expressed genes by the subgroup within the non-classical monocyte subset identified on the PCA score plot. The subgroup is marked by filled blue pluses in the PCA score plot.</p

Multimodal expression in the three monocyte subsets.

<p><b>Bold</b> = Multimodal expression <i>P</i> <0.05, <i>Italic</i> = Unimodal expression</p><p>Multimodal expression in the three monocyte subsets.</p

Multimodal variation in expression levels across the three monocyte subsets.

<p>Violin plot demonstrating multimodal variation in gene expression levels of the 85 genes examined in the monocyte subsets. The classical monocytes, intermediate monocytes and non-classical monocytes are indicated in the figure by red, green and blue, respectively. The data depict the multimodal expression levels of the genes listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0144351#pone.0144351.t004" target="_blank">Table 4</a> calculated by using the Hartigans dip test (<i>P <</i> 0.05).</p

Single-cell gene expression analysis on human monocytes.

<p>Human monocytes were single-cell sorted according to the expression of the cell surface markers CD14 and CD16 and gene expression on single cells was assessed. A) Representative plot of flow cytometry analysis demonstrating the monocyte subset gating strategy, used to single-cell sort monocytes from the three classified monocyte subsets (<i>n =</i> 1). Classical (CD14⁺⁺CD16^-) monocytes, intermediate (CD14⁺⁺CD16⁺) monocytes and non-classical (CD14⁺⁺CD16⁺) monocytes are depicted in the upper left quadrant, upper right quadrant and lower right quadrant, respectively. B) Principal component analyses (PCA) of single-cell PCR gene expression analysis data showing genetic clustering of the three monocyte subsets. The PCA plot confirmed the classification of the three human monocyte subsets done by flow cytometry, visualized by gene families. Each dot represents a single cell. C) Heatmap of gene expression values for PCA showing hierarchical clustering of single-cell PCR gene expression data from the three human monocyte sub-populations. The analysis revealed cellular heterogeneity by distinct gene signatures. Red circles = classical monocytes (<i>n</i> = 94 cells), green triangles = intermediate monocytes (<i>n</i> = 92 cells), and blue pluses = non-classical monocytes (<i>n</i> = 80 cells).</p

Details of RABV samples investigated in this study.

<p>Details of RABV samples investigated in this study.</p

Comparison of substitution rates per site of RABVs.

<p>Comparison of substitution rates per site of RABVs.</p

Phylogram of 49 arctic foxes generated from concatenated sequences of 12 mitochondrial protein coding sequences (ATP6, ATP8, COX1, COX2, COX3, CYTB, ND1, ND2, ND3, ND4, ND4L and NDS).

<p>Bootstrap values higher than 40% are indicated. The minimum identity of sequences to be grouped into a fox cluster was set to 99.7%.</p

Maximum clade credibility (MCC) phylogenetic tree using complete genome sequences from 79 RABV samples sequenced in this study (Table 1).

<p>The minimum identity of sequences to be grouped into a single subclade within arctic lineage 3 was set to 99.5%.</p