title: GSM8978557: Mice model of SLE treated with JPZS-1; Mus musculus; Bisulfite-Seq
library_construction_protocol: DNA was extracted from CD4+ T cells using a TIANamp Genomic DNA Kit (Tiangen Biotech Co. Ltd., Beijing, China, cat. DP304) The methylation level of the Cytosine-phosphate-guanine (CpG) site within the transcription start site region of the Foxp3 gene was evaluated utilizing the MethylTarget™ assays (targeted bisulfite sequencing) developed by Genesky BioTech (Shanghai, China). Genomic DNA was extracted and subsequently subjected to methylation using the EZ DNA Methylation Gold Sulfurous Acid Kit (ZYMO, CA, USA) according to the manufacturer's instructions. Primers for the bisulfite-converted DNA samples were designed using FastTarget V4.1 (Genesky, Shanghai, China). The specific primer sequences (5′-3′) were as follows: Foxp3-(Treg-specific demethylated regions) TSDR: forward: TTGGGTTTTTTTGGTATTTAAGA, reverse: AAATCTACATCTAAACCCTATTATCACAA. Following the bisulfite conversion of genomic DNA, PCR was conducted to amplify the target DNA sequences. The amplified products were sequenced using an Illumina HiSeq sequencer (Illumina, CA, USA).