GSM3823946: RNA_Treg_Tfam_Ctrl_R2; Mus musculus; RNA-Seq
[RNA-seq] Total RNA was extracted from about 1×10e6 sorted YFP+ Tregs pooled from spleen and lymph node of 2 to 3 Tfamfl/+Foxp3Yfp-Cre/+ or Tfamfl/flFoxp3Yfp-Cre/+ female littermate mice with RNAeasy Mini Kit (Qiagen), followed by DNase treatment (TURTBO DNA-freeTM Kit). [ATAC-seq] 5x10e4 sorted splenic Tregs were lysed in lysis buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% IGEPAL CA-630). Pelleted nuclei were resuspended in transposition reaction mix with Tn5 transposase (Illumina) and incubated at 37°C for 30 min. [RNA-seq] mRNA was purified with NEBNext Poly(A) mRNA Magnetic Isolation kit (NEB) and RNA-seq libraries were generated using NEBNext Ultra II RNA library Prep Kit (NEB). [ATAC-seq] The DNA was purified with MinElute PCR Purification Kit (Qiagen) and was used as a template of PCR with 10 cycles of amplification. The library was cleaned up with 1.2x SPRIselect beads (Beckman Coulter) before sequencing.
GEO Accession
GSM3823946