biostudies-arrayexpressAzusa TerasakiMus musculushttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-16050Analysis of total RNA of B16 melanoma cell line after co-culture with mitochondria-tagged (Dendra2 PhAMexcised mitochondria reporter - also known as mtD2, strain no. 018397mtD2) leukocytes. The cells were sorted to two groups: mtD2+ B16 and mtD2- B16 cells.biostudies-arrayexpressNucleic Acid Extraction - Total RNA was isolated using Qiagen RNEasy Plus isolation kit.Sequencing - Bulk RNAseq was performed using the DNBSEQ platform.Sample Collection - After tumor-immune co-cultures, cells were resuspended in FACS buffer (PBS in 0.5% BSA) and stained with DAPI and CD45. Live tumor cells (TdTomato+CD45-DAPI-) were sorted into Dendra2+ and Dendra2- subpopulations and pelleted immediately.Library Construction - 1) mRNA enrichment and purification: Oligo dT Selection (mRNA enrichment): Use oligo dT beads to enrich mRNA with poly A tail. 2) RNA fragment and reverse transcription Fragment the RNA and first-strand cDNA was generated using random N6-primed reverse transcription, followed by a second-strand cDNA synthesis with dUTP instead of dTTP. 3) End repair, add A and adaptor ligation The synthesized cDNA was subjected to end-repair and then was 3' adenylated. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments; 4) PCR amplification Prior to PCR amplification, the dUTP-marked strand is selectively degraded by Uracil-DNA-Glycosylase(UDG). The remaining strand is amplified to generate a cDNA library suitable for sequencing. Many rounds of PCR amplification were performed to enrich the purified cDNA template using PCR primer; 5) Single strand separation and cyclization Denature the PCR product by heat and the single strand DNA is cyclized by splint oligo and DNA ligase; 6) DNA nanoball synthesisOrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - The raw RNA sequencing data was mapped to mouse reference genome hg38 using the STAR aligner, and genes annotated in Gencode v3664 was quantified using featurecounts in the subread package.MetabolomicsUnknownTranscriptomicsGenomicsProteomicsDNBSEQ-T7RNA-seq of coding RNAMus musculusAzusa TerasakifalseMitochondria transfer from immune cells enables lymph node metastasisAnalysis of total RNA of B16 melanoma cell line after co-culture with mitochondria-tagged (Dendra2 PhAMexcised mitochondria reporter - also known as mtD2, strain no. 018397mtD2) leukocytes. The cells were sorted to two groups: mtD2+ B16 and mtD2- B16 cells.2025-12-25T00:00:00Z2026-05-30T16:00:21.016Z2025-11-13T15:17:19.756ZE-MTAB-16050ERP183980EFO_0002944EFO_0004170EFO_0005518EFO_0003816EFO_0003738EFO_0004184