GSM5531491: H3K9me3 Fzr1-OE-input DNA; Mus musculus; ChIP-Seq
KAPA Hyper Prep Kit was used for cytoplasmic lysis of the indicated samples. For RNA-seq, reverse transcription was performed directly with the cytoplasmic lysates of the indicated samples. A total cDNA library was then amplified over 18-20 cycles for library construction. The amplified cDNA was fragmented, and then a KAPA Hyper Prep Kit was used to generate sequence libraries. Paired-end 125 bp or 150 bp sequencing was further performed on a HiSeq 2500 system (Illumina) at Novogene. For H3K9me3 ChIP-Seq, 500 cells were used per reaction, and two replicates were performed. All isolated cells were washed three times in 0.5% BSA in PBS solution (Sigma) to avoid possible contamination. The NChIP procedure was performed as previously described (Brind'Amour et al, 2015). Histone H3K9me3 Antibody (39161, Active Motif) was used for each immunoprecipitation reaction. The sequence libraries were generated using the KAPA Hyper Prep Kit for the Illumina platform (kk8504), following the manufacturer's instructions. Paired-end 125-bp or 150-bp sequencing was performed on a HiSeq 2500 or X10 (Illumina) at Berry Genomics. For RRBS, the subcutaneous adipose tissues of three 4-month-old mice in each group were collected, cryopreserved at -80°C. The whole-genomic DNA extracted from tissue was fragmented by ultrasound, A bases were added for end-repair, and methylated adaptor fragments were added. The obtained DNA was subjected to bisulfite treatment, and the final DNA library was obtained after PCR amplification. Paired-end 150 bp sequencing was further performed on a HiSeq 2500 system (Illumina) at Novogene.