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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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MiCLIP profiling of m6A sites in human CD8⁺ T cells during activation, with Input and noUV controls

ABSTRACT: Methylation individual-nucleotide-resolution crosslinking and immunoprecipitation (miCLIP) was performed in human primary CD8⁺ T cells subjected to in vitro activation with αCD3/αCD28 beads. To map m⁶A sites and assess their dynamic signatures during T cell activation, we applied an improved iCLIP (iiCLIP) protocol using an anti-m⁶A antibody (Abcam) on 750–100 ng of polyadenylated RNA. Samples were collected from non-activated (NoAct), Day 1-activated, and Day 5-activated CD8⁺ T cells isolated from healthy donors (NoAct: 4 donors; Day 1: 5 donors; Day 5: 4 donors). Two experimental controls were included: (i) “Input” libraries prepared without the m⁶A immunoprecipitation step (4 donors each for NoAct, Day 1, and Day 5), and (ii) “noUV” libraries generated without UV crosslinking (3 donors total).

INSTRUMENT(S): Illumina HiSeq 4000

ORGANISM(S): Homo sapiens

SUBMITTER: Jernej Ule

PROVIDER: E-MTAB-15643 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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