Region specific astrocyte response to mutant GFAP in Alexander disease model mice during postnatal development.

Gene expression analysis was performed in Gfap+/+, Gfap+/R236H, and mGFAPTg170-2 transgenic mice, using the Aldh1l1-eGFP-L10a transgenic line (JD130) to isolate translating ribosomes and purify astrocyte specific transcripts for RNAseq. Hippocampus and corpus callosum were collected from male mice at P14 in an FVBB6F1 background. Translating ribosomes were isolated using antibodies against EGFP to precipitate the EGFP-L10a fusion protein and the associated RNA purified for sequencing and gene expression analysis. N = 3 for each genotype with each sample consisting of tissues pooled from 5 mice (15 mice total).本研究针对Gfap+/+、Gfap+/R236H及mGFAPTg170-2转基因小鼠开展基因表达分析，使用Aldh1l1-eGFP-L10a转基因小鼠品系（JD130）分离翻译核糖体，并纯化星形胶质细胞特异性转录本以进行RNA测序（RNAseq）。实验采集FVBB6F1背景下雄性小鼠在出生后第14天（P14）的海马体与胼胝体组织。通过抗EGFP抗体沉淀EGFP-L10a融合蛋白及其结合的RNA，对纯化得到的RNA开展测序与基因表达分析。每种基因型设置3个生物学重复，每个样本由5只小鼠的组织混合制备，本实验总计使用15只小鼠。