AR in skeletal muscle

Determine AR cistrome in skeletal muscle and correlate it with the presence of H3K4me2. Overall design: Muscle nuclei were extracted as described (Joshi et al., 2016). After sonication, chromatin was immunoprecipitated with anti-AR and anti-H3K4me2 antibodies. sequenced with an Illumina Hiseq 4000 as paired-end 100 bp reads, and mapped to the mm10 reference genome using Bowtie 2 (Langmead and Salzberg, 2012). Uniquely mapped reads were retained for further analysis. MACS2 algorithm (https://github.com/taoliu/MACS/) (Zhang et al., 2008) was used for peak calling with non-immunoprecipitated chromatin as control.本研究旨在鉴定骨骼肌中的雄激素受体（Androgen Receptor, AR）顺式调控组（cistrome），并将其与组蛋白H3赖氨酸4二甲基化（H3K4me2）的存在状态进行关联分析。实验设计概述：按照文献（Joshi等，2016）所述方法提取骨骼肌细胞核。经超声破碎处理后，使用抗AR抗体与抗H3K4me2抗体对染色质进行免疫沉淀。随后采用Illumina HiSeq 4000平台进行双端100 bp读长测序，并利用Bowtie2（Langmead与Salzberg，2012）将测序读段（reads）比对至mm10参考基因组。仅保留唯一比对的读段（reads）用于后续分析。采用MACS2算法（https://github.com/taoliu/MACS/，Zhang等，2008）以未进行免疫沉淀的染色质作为对照进行峰调用。