Figure 4.

Effect of miR-125b mimics and inhibitors on MK differentiation of human umbilical CB MNCs. Cells were cultured in a megakaryocytic differentiation medium for 6 days, and transiently transfected with miR-125b mimics or inhibitors at day 1. (a) Relative miR-125b expression analyzed by qPCR. miR-125b expression is normalized to U6. (b) Overexpression of miR-125b in MNCs induces the expression of the MK integrins CD41 and CD61. (c) Relative expression of MK marker genes. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a housekeeping control in the experiment. qPCR results from cells transfected with NC mimics are assigned an arbitrary value of 1. (d) Transient transfection of the designated inhibitor effectively downregulates the expression of miR-125b. (e) Downregulated miR-125b expression in MNCs reduces the expression of the MK integrins CD41 and CD61. (f) Relative expression of MK marker genes. GAPDH was used as a housekeeping control in the experiment. qPCR results from cells transfected with NC inhibitor mimics are assigned an arbitrary value of 1. Cells were obtained from four different donors and each sample was tested with three independent experiments for data presentation. All of the data are expressed as mean±S.D. Student's t-tests were used for statistical analysis. *P<0.05 and **P<0.01