biostudies-arrayexpressNeil MorganMus musculushttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15572To identify the mechanism through which SLFN14 may regulate megakaryopoiesis and platelet function, we performed RNA sequencing analysis to define alterations in gene activity arising as a result of conditionally knocking down Slfn14 in vivo. Bulk RNA-sequencing was performed on total RNA extracted from mature murine Slfn14fl/flPF4-Cre derived MKs and peripheral blood platelets and compared to Slfn14+/+PF4-Cre control MKs/platelets. All individual samples were run in triplicate and normalised before comparisons between each SLFN14 mutant.biostudies-arrayexpressLibrary Construction - Illumina/NEB kit protocol - Libraries were prepared using the NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina with NEBNext® rRNA Depletion Kit v2 (Human/Mouse/Rat) according to the manufacturer’s protocol. During this process, the libraires were indexed using NEBNext® Multiplex Oligos for Illumina® (96 Unique Dual Index Primer Pairs) Set 4. The prepared libraries were quantified via a fluorometric method involving a Promega QuantiFluor dsDNA assay and qualified using electrophoretic separation on the Agilent TapeStation 4200.Sample Collection - Mice were exsanguinated under terminal anesthesia by isoflurane/O2 (5%) gas. Blood was drawn from the inferior vena cava, using a 25-gauge needle, into 1:10 (volume-to-volume ratio) acid citrate dextrose anticoagulant.Sequencing - Sequencing was performed using the Illumina HiSeq 4000.Nucleic Acid Extraction - Platelets and megakaryocytes were isolated and RNA was extracted using the RNeasy mini kit (Qiagen) according to manufacturer’s instructionsOrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - RNA sequencing performed on platelet and MK RNA in triplicate groups from individual mice (Slfn14fl/flPF4-Cre, Slfn14f+/+PF4-Cre) and normalised comparing mutant and WT groupsMetabolomicsUnknownTranscriptomicsGenomicsProteomicsIllumina HiSeq 4000RNA-seq of total RNAMus musculusNeil MorganfalseRNA-seq shows genes enhanced in translation and transcription pathways in MKs and platelets from Slfn14fl/flPF4-Cre mouse modelTo identify the mechanism through which SLFN14 may regulate megakaryopoiesis and platelet function, we performed RNA sequencing analysis to define alterations in gene activity arising as a result of conditionally knocking down Slfn14 in vivo. Bulk RNA-sequencing was performed on total RNA extracted from mature murine Slfn14fl/flPF4-Cre derived MKs and peripheral blood platelets and compared to Slfn14+/+PF4-Cre control MKs/platelets. All individual samples were run in triplicate and normalised before comparisons between each SLFN14 mutant.2025-09-29T00:00:00Z2026-05-28T10:15:20.661Z2025-09-12T14:40:47.614ZE-MTAB-15572ERP180028EFO_0002944EFO_0004170EFO_0009653EFO_0005518EFO_0003816EFO_0004184