Fig. 4

Functional consequences of miR-10a overexpression in hematopoietic stem progenitor cells. CD34⁺ primary hematopoietic cells purified from cord blood were transduced with a lentivirus expressing the pri-miR-10a precursor or the empty vector and grown in complete medium containing SCF, TPO and Flt3-L. a Efficient overexpression of miR-10a was assessed by qRT-PCR on CD34⁺ cells 4 days after transduction. Results are expressed as the expression of miR-10a in one representative experiment, normalized on RNU6–1 expression and compared to empty vector condition (n = 2). b Cells were counted at different times after transduction. Results are expressed as fold increase after stimulation. The graph represents the mean values of 2 independent experiments. c After 7 days of culture, differentiation markers expression was assessed by flow cytometry. The graph represents the percentage of positive cells in one representative experiment performed in duplicate (n = 2) (d). After 7 days of liquid culture, cells were implanted in semi-solid medium in presence of EPO (for erythroid colonies) or G-CSF (for myeloid colonies). The graph represents the mean colony number for 1000 cells seeded obtained in 2 independent experiments. e Stem cell renewal was assessed by culturing transduced normal CD34⁺ cells on MS5 stromal cells for 5 weeks. Clonogenic capacity was then assessed in semi-solid medium in presence of EPO (for erythroid colonies) or G-CSF (for myeloid colonies). Colonies were counted after 2 weeks. Graph shows mean colony number for 1000 cells seeded of 4 independent experiments