PU.1 induces a specific miR profile. PUER and PU.1 KO cells were treated with solvent or with OHT (100nM) for 18 hours and profiled for miR expression. Monocytic differentiation of PUER was controlled by FACS analysis showing induction of CD11b and by phenotype (supplemental Figure 1). (A) Strategy of PU.1-regulated miR identification. Raw data of the PUER and PU.1 KO cells were normalized to global mean. PU.1-specific candidates were then identified using SAM for PUER cells (18 hours for solvent vs 18 hours for OHT) using a 2-fold change for up-regulated and a 0.75-fold change for down-regulated miRs as a cutoff. To control for unspecific OHT effects on miR expression, those miR that were also differentially expressed in PU.1 KO cells after 18 hours of OHT treatment compared with the solvent control were excluded from further analyses. (B) SAM plot of the miR expression differences between OHT and mock-treated PUER after exclusion of the miR expression changes resulting from OHT treatment of PU.1 KO cells. Selected PU.1-regulated miRs are indicated (arrow); all PU.1-regulated miRs are listed in Table 1. Arrays were performed in biologic triplicate for PUER cells and in duplicate for PU.1 KO cells.