miRNA expression in LT-HSCs upon Thrombopoietin stimulation via STAT5

<strong>File description:<br></strong> <strong><br></strong> <strong>MX: STAT5-deficient</strong> <strong>FL: STAT5-wt</strong> <strong>1/2: cohort 1 and 2</strong> <strong>+: TPO-stimulated</strong> <strong>-: unstimulated<br></strong> <strong><br></strong> <strong>MiRNA expression in murine long-term repopulating hematopoietic stem cells upon Thrombopoietin stimuation via STAT5A/B</strong> Conditional deletion of STAT5A/B was induced in 6-week old mice (STAT5fl/fl x Mx1:Cre and STAT5fl/fl; 16-20 mice per group and experiment) via Poly(I:C) injections as previously described. LT-HSCs were isolated from 12-16 week old mice via FACS sorting. 5000-10000 viable LT-HSCs were starved for 5h and subsequently stimulated wit 100ng/ml TPO (Peprotech) for 2h. The RNA was isolated by using a miRNeasy mini kit (Qiagen). cDNA was synthesised from total RNA using a microRNA gene-specific RT-primer pool according to the MicroRNA Megaplex Assay protocol (Megaplex™ Pools - Applied Biosystems). The preamplification reaction was performed according to the manufacture´s protocol. The TaqMan Array MicroRNA Rodent Cards were analysed using the 384 well TaqMan Low Density Array default thermal cycling conditions (ABI PRISM 7900HT).