biostudies-arrayexpressRong HuaMus musculushttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-14988Using isolated and purified round and elongated spermatids from the testes, Ribosome sequencing was performed to assess the impact of MEX3D protein deficiency on RNA metabolism in spermiogenesis.biostudies-arrayexpressSequencing - Libraries were sequenced on a PE150 run on Illumina Novaseq X Plus instrument.Library Construction - A fraction of RNAs (10-200 ng) were used for cDNA library construction. After 3' polyadenylation, RNA was ligated to a linker at room temperature and purified using streptavidin magnetic beads. cDNA libraries were then generated through reverse transcription and PCR amplification, with no more than 14 cycles of PCR performed per group.Nucleic Acid Extraction - Snap-frozen spermatids were cryoground into powder on dry ice, and then resuspended in ice-cold lysis buffer. After digest with RNase I, RNA was purified for ribosome-protected small RNA fragments via a 15% polyacrylamide TBE-urea gel. The gel debris was removed using a Spin-X column (Corning, 2628866), followed by ethanol precipitation. Purified RNA fragments (26-32 nucleotides) were resuspended in nuclease-free water and quantified using Qubit 2.0 Fluorometer (Invitrogen).Sample Collection - Round and elongated spermatids were isolated and purified from adult mouse testes.OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - Gene-level quantification was performed using featureCounts (v2.0.6), counting paired-end reads (`-p --countReadPairs`) aligned to exon features (`-t exon`) and assigning counts to genes (`-g gene_id`).Sequence Alignment - For data analysis, paired-end reads were quality-trimmed using fastp (v0.23.4) with default parameters and 16 processing threads. Clean reads were aligned to the mouse reference genome (GRCm39, Ensembl release 112) using STAR (v2.7.11b). The resulting alignments were sorted by coordinates and stored in BAM format.UnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq XRibo-seqMus musculusZerui WangRong HuafalseRibosome Sequencing of Testicular Spermatids in Mex3d Knockout and Wild-Type MiceUsing isolated and purified round and elongated spermatids from the testes, Ribosome sequencing was performed to assess the impact of MEX3D protein deficiency on RNA metabolism in spermiogenesis.2026-03-01T00:00:00Z2026-03-01T02:02:46.699Z2025-03-31T12:58:52.877ZE-MTAB-14988ERP171026E-MTAB-14966EFO_0002944EFO_0004170EFO_0004917EFO_0008891EFO_0005518EFO_0003816EFO_0004184