Comparison of gene expression profiles of miR-142-/- primary megakaryocytes and WT primary megakaryocytes. Mus musculus

Whole fetal livers were collected from mouse fetuses at embryonic day 14.5 (E14.5), and single-cell suspensions were prepared by successive passage through 18-, 21 and 23-gauge needles. Fetal liver cells were maintained in Dulbecco modified Eagle medium (DMEM; Invitrogen) supplemented with 10% fetal bovine serum (FBS; Invitrogen), 100 U/ml penicillin, 100µg/ml streptomycin, and 50ng/ml recombinant human thrombopoietin (TPO; Peprotech). After 5 days of culture, megakaryocytes were purified using a discontinuous bovine serum albumin gradient (BSA, Sigma–Aldrich; 3%, 1.5%, and 0%). Total RNA was isolated with Tri–Reagent (MRC) following manufacturer’s instructions, and its quality was assessed with ND–1000 Nanodrop (Peqlab) and on a 1.5% agarose gel. Overall design: Gene expression was measured in primary megakaryocytes cultured from miR-142-/- fetal livers and wild type fetal livers at E14.5. Two independent experiments were performed.