GSM4072391: AhrChIP_iTreg_WT_R2; Mus musculus; ChIP-Seq
Th17 and iTreg were differentiated from sorted naïve CD4+ T cells as described above. For ChIP of Ahr, cells were treated with FICZ (200 nM) for 4 hours before harvest. Cells were cross-linked with 1% formaldehyde for 15 min. Chromatin was sheared by sonication with Bioruptor Pico (30” on and 30” off for 25 cycles) and immunoprecipitated with anti-Ahr (Enzo Life Science), anti-RORγt (eBioscience, AFKJS-9) or anti-Foxp3 (Abcam, ab150743) using iDeal ChIP-Seq Kit for transcription factors or True MicroChIP kit (Diagenode). [ChIP-seq] Eluted DNA was made into indexed library according to the manual of NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB). The library was cleaned up with 1.2x SPRIselect beads (Beckman Coulter) before sequencing.