Expression of miR-139-5p suppresses proliferation in hematopoietic progenitors and CML cells. (A) Cell cycle profile was analyzed using Ki67/DAPI staining in Kcl22 cells after transfection with empty or miR-139-5p–expessing vectors (N = 3). (B) Cell cycle progression was analyzed using PI staining in K562 cells after transfection with empty or miR-139-5p–expessing vectors (N = 5). (C) Expression of genes that are associated with cell survival and apoptosis was analyzed in miR-139-5p–transfected K562 cells. Results are presented relative to those of cells transfected with control empty vector (N = 3). (D) Expression of cell cycle–related genes was analyzed in miR-139-5p–transfected K562 cells. Results are presented relative to those of cells transfected with control empty vector (N = 3). (E) Expression of p27, p21, and miR-139-5p in K562 cells after transfection with empty or miR-139-5p–expressing vectors was measured using quantitative RT-PCR analysis (N = 3). (F) Cell proliferation was analyzed in LSK (Lin⁻c-Kit⁺Sca-1⁺) of mice reconstituted with miR-139-5p-RV. Bar graph represents mean ± SEM (N = 3). (G) Cell proliferation was analyzed in LSK cells of mice reconstituted with miR-139T-RV. Bar graph represents mean ± SEM (N = 3). (H) Expression of cyclin D1, cyclin D3, and p27 was measured in LSK cells purified from mice reconstituted with miR-139-5p-RV (N = 3). (I) Representative cytometric analysis of granulocytes (left) and myeloid progenitors, LK population (middle) from reconstituted mice as described in the figure. Upper panels, empty-NGFR and empty-GFP; middle panels, p210-NGFR and empty-GFP; lower panels, p210-NGFR and miR-139-5p-GFP. The mean frequency of granulocytes and LK population is depicted (right, mean ± SEM). (J) Expression of miR-139-5p and p210 fusion transcript was analyzed in Lin⁻c-Kit⁺ HSPCs purified from reconstituted mice using quantitative RT-PCR. Data represent the mean ± SEM (N = 4).