CD8/CD4 ratio in PBMCs from young and old marmosets.

*<p>Only FACS analysis, but not qPCR, was done with PBMCs from these three marmosets.</p

Sequences of qPCR primers for CD markers and cytokines.

a)<p>Hyphen indicates a nucleotide identical to human sequences.</p>b)<p>Dot indicates a shift nucleotide to marmoset sequences.</p

The expression ratios of CD8 to CD4 (CD8∶CD4) and Th1-related genes to Th2-related genes.

<p>The ratio of CD8∶CD4 (left panel), IFN-γ∶IL-4 (middle panel) and IL-2∶IL-4 (right panel) in human and common marmoset leukocytes, spleen, lymph node and thymus are shown. Significant differences in the CD8∶CD4, IFN-γ∶IL-4 and IL-2∶IL-4 ratios were found between human leukocytes and common marmoset tissues (*<i>P</i><0.05).</p

The expression levels of CD antigens and cytokine genes in common marmoset and human leukocytes.

<p>The expression level of each gene is shown as a logarithmic histogram of absolute copy numbers per µg of total RNA. Means and standard deviations of eight individuals are indicated. Asterisk indicates statistically significant differences between marmosets and humans by Student's <i>t</i>-test (*<i>P</i> value<0.05, **<i>P</i> value<0.01).</p

Absolute copy numbers of candidate reference genes.

<p>The expression level of each gene in 13 tissues is shown as a logarithmic histogram of absolute copy numbers per µg of total RNA. Means and standard deviations of four individuals are indicated. GAPDH: glyceraldehyde-3-phosphate dehydrogenase; ACTB: actin, beta; rRNA: 18S ribosomal RNA; B2M: beta-2-microglobulin; UBC: ubiquitin C; HPRT: hypoxanthine phosphoribosyltransferase 1; SDHA: succinate dehydrogenase complex, subunit A; TBP: TATA-box binding protein.</p

Gene expression stability and pairwise variation of candidate reference genes using <i>geNorm</i> analysis.

<p>(A) and (B): Average gene expression stability values M of the remaining reference genes during stepwise exclusion of the least stable gene in the different tissue panels are shown. Data are divided into two figures to avoid closely-packed lines. See also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056296#pone-0056296-g003" target="_blank">figure 3</a> for the ranking of genes according to their expression stability. (C) Pairwise variation analysis was used to determine the optimal number of reference genes for use in qPCR data normalization. The recommended limit for V value is 0.15, the point at which it is unnecessary to include additional genes in a normalization strategy.</p

The ratio of CD4⁺ to CD8⁺ cells in common marmoset and human peripheral blood mononuclear cells (PBMCs) by flow cytometry.

<p>Representative scattered plots of FSC and SSC are shown in the left panels. Middle panels represent a histogram of CD3 analyzed in the lymphocyte gate. Gated CD3⁺ cells were analyzed for CD4 and CD8 expression (right panels).</p

Ranking of gene expression stability of candidate reference genes using <i>geNorm</i> analysis.

<p>Candidate reference genes are ranked in order of stability for each tissue with the two most stable genes at the left and the least stable at the right.</p