Identifying novel miRNAs through high-throughput sequencing. (A) Distribution of known and candidate novel miRNAs in normal and malignant B cells. Samples are listed in the same order as Table 1. (B) Distribution of known miRNA expression among the normal B-cell subsets. The majority of miRNAs are shared between the B-cell types. (C) Distribution of candidate novel miRNA expression among the normal B-cell subsets. The majority of miRNAs are shared between the B-cell types. (D) Proportion of samples in which candidate novel miRNAs were identified. The majority of miRNAs are expressed in more than one sample. (E) Proportion of candidate novel miRNAs identified in 20 additional sequencing datasets. (F) Conservation of novel human miRNAs in chimpanzee, rhesus monkey, mouse, and dog. Ninety percent of the 286 novel miRNAs were found to be conserved across humans and one or more additional species, and more than 80% were found to be conserved across humans and 2 or more additional species. (G) Validation of 86 novel miRNAs candidates by an alternative method of measurement, real-time PCR. Maximum measured values for miRNAs expressed in at least 10% of the samples are shown in log2-scale. Detection threshold is the expression level corresponding to C_T ≤ 35. (H) miRNA profiling distinguishes the molecular subgroups of DLCBL that were defined based on gene expression profiles. ABC refers to activated B-cell–like DLBCL. GCB refers to germinal center B-cell–like DLCBL. Unclassified cases are those that did not meet criteria for either group. Black bars indicate candidate novel miRNAs that were differentially expressed between the 2 groups.