MDDCs exhibit unique stage-specific patterns in miRNA expression. (A) Flow cytometric analysis showing cell surface phenotype during MDDC differentiation. Monocytes and DCs can be distinguished by their CD14 and DC-SIGN expression profiles. Data from one representative donor (of 3) are shown. (B) Quantification of DC-SIGN/CD14 expression ratios in 5 independent donors from 7 independent experiments. Ratios at day 0 were measured in 4 of 7 independent experiments. Ratios at days 1, 3, and 5 were measured in 7 of 7 independent experiments. Ratios were calculated using MFI of DC-SIGN/ MFI of CD14 and normalized to the day 5 ratio of each donor, which was set at 1.0. (C) PCA of all 389 miRNA probes shows distinct clustering of expression profiles from day 0, 1, 3, and 5 differentiated monocytes. First and second principal components, represented on the x-axis and y-axis, respectively, account for approximately 32% and 12% of the variability between the various samples. Different time points (in 3 different donors; each represented by one circle) are represented by the indicated colors. (D) Hierarchical clustering of statistically significant differential miRNAs (human miRNAs) with analysis of variance (P < .005). Boldface type represents upregulated miRNAs. Downregulated miRNAs are underlined. Remaining miRNAs have sinusoidal expression. Clustering on the top represents the relationships between the various time points, and each column in a given day represents an independent donor. Color legend indicates relative log scale intensity of expression.