Transcriptome profiling of purified mouse platelets from Nxf1 mutant and control mice [Platelet]

Purpose: Nxf1 is thought to be an essential nuclear exporter of messenger RNA (mRNA) in eukaryotic cells. Whether perturbations in the Nxf1 pathway affect mammalian physiology is not known. The aim of this study is to determine the impact of a Nxf1 mutation in the representation of mRNA transcripts in platelets. Methods: Platelets were purified from individual males. Blood was obtained by cardiac puncture into 0.1 volume of Aster Jandl citrate-based anticoagulant. Mouse platelet rich fraction was obtained by centrifugation of the murine blood at 125 g for 8 min at room temperature, followed by centrifugation of the supernatant buffy coat at 125 g for 8 min. Mouse platelets were washed by two sequential centrifugations at 860 g for 5 min in 140 mM NaCl, 5 mM KCl, 12 mM trisodium citrate, 10 mM glucose, and 12.5 mM sucrose, pH 6.0.The platelet pellet was resuspended in 10 mM Hepes, 140 Mm NaCl, 3 mM KCl, 0.5 mM MgCl2, 10 mM glucose, and 0.5 mM NaHCO3, pH 7.4. The purity of each platelet suspension was assessed by flow cytometry and suspensions for which more than 98% of total events were CD41+ platelets were pooled together. RNA was purified with Norgen cytoplasmic and nuclear fractionation RNA purification kit. Platelets were purified from individual Nxf1 mutant males (littermate controls are available from GEO accession GSE75896). The purity of each suspension was assessed by flow cytometry. Suspensions of identical genotype for which more than 98% of total events were CD41+ platelets were pooled together. Pooled platelet preparations from mutant (this study) and wild-type littermate controls (GSE75896) were prepared on 3 distinct days. The GSM4196370-GSM4196372 sample records are duplicated sample records of GSM1969530-GSM1969532 (in GSE75896) for the convenient retrieval of the complete raw data from SRA.