Determination of Lsd1 function in inguinal white adipose tissue (ingWAT) by global transcriptome analysis

To investigate the role of Lsd1 in ingWAT we crossed mice harboring conditional Lsd1 alleles (Zhu et al., 2014) with the Adipoq-Cre deleter strain (Eguchi et al., 2011), which results in Cre-mediated loss of Lsd1 selectively in adipocytes. Transciptome analysis of ingWAT obtained from control and knock-out mice revealed 727 genes differentially expressed between Lsd1 knock-out and control mice at 6 weeks of age (p-value<10-2). We selectively knocked-out Lsd1 in adipose tissue by crossing Lsd1 floxed mice to Adipoq-Cre mice. RNA from inWAT of 6 week.old control (Lsd1 floxed mice) and knock-out mice was extracted with trizol according to the manufacturer protocol. RNA samples were sequenced by the standard Illumina protocol to create raw sequence files (.fastq files). We annotated these reads to the mm10 build of the mouse genome using TopHat version 2. The aligned reads were counted with the homer software (analyze RNA) and DEG's were identified using EdgeR and DESeq version 1.8.3.为探究Lsd1在腹股沟白色脂肪组织（inguinal white adipose tissue, ingWAT）中的功能，我们将携带条件性Lsd1等位基因（Zhu等，2014）的小鼠与Adipoq-Cre敲除工具株（Eguchi等，2011）进行杂交，该策略可实现脂肪细胞中Cre介导的Lsd1特异性缺失。对对照小鼠与Lsd1敲除小鼠的腹股沟白色脂肪组织进行转录组分析，结果显示在6周龄时，两组小鼠间存在727个差异表达基因（p值<10⁻²）。我们通过将携带Lsd1 floxed（两侧带有LoxP重组位点）等位基因的小鼠与Adipoq-Cre小鼠杂交，实现了脂肪组织中Lsd1的特异性敲除。采用Trizol试剂（按试剂盒制造商说明书操作）提取6周龄对照小鼠（Lsd1 floxed小鼠）与敲除小鼠的腹股沟白色脂肪组织总RNA。按照标准Illumina测序流程对RNA样本进行测序，得到原始测序文件（.fastq格式文件）。我们使用TopHat v2软件将这些测序读段比对到小鼠基因组mm10组装版本。随后使用Homer软件的analyze RNA模块对比对后的读段进行计数，并通过EdgeR与DESeq v1.8.3软件鉴定差异表达基因（differentially expressed genes, DEGs）。