m6A modification in effector CD4+ T cells [m6A-miCLIP-seq]

GP61-primed effector CD4+ T cells were isolated from Ctrl or Mettl3-deficient SMARTA mice. Total RNAs were extracted with TRIzol reagent, and mRNAs were then isolated with Dynabeads® mRNA purification kit, followed by stardard m6A-miCLIP-SMARTer-seq with some modifications. Raw sequencing reads were aligned to the mouse genome (mm10) with BWA, and then m6A sites were determined. Overall design: Ctrl or Mettl3fl/flCd4-Cre SMARTA mice were primed with LCMV GP61 peptide , and 16-18 h later, CD62LloCD44hiCD4+ T cells were sorted and subjected to m6A-miCLIP-SMARTer-seq.