Gene expression differences between TAF4b-KO, Het and WT ovaries at 3 weeks of age

The rapid decline of ovarian function in TAF4b-null mice begins in early postnatal life and follicle depletion is completed by sixteen weeks. To uncover differences in gene expression that may underlie accelerated ovarian aging, we compared genome-wide expression profiles of three week old, pre-pubescent TAF4b-null and wild-type ovaries. Total RNA from 9, 3-week-old mice (3 wild-type, 3 TAF4b-heterozygous, 3 TAF4b-null) was obtained as described as above. RNA quality was checked using a Bioanalyzer, and concentration determined using a Nanodrop. 300 ng of each RNA sample was used in the Affymetrix Whole-transcript Sense Target Labeling Assay (Rev 3) followed by hybridization to a GeneChip Mouse Gene 1.0 ST Array. 9 GeneChips were used to provide biological triplicates of each genotype. The Affymetrix Expression Console (v 1.1) was used to normalize data and determine signal intensity (RMA-Sketch).TAF4b敲除（TAF4b-null）小鼠的卵巢功能快速衰退始于产后早期，且卵泡耗竭于16周龄时完全完成。为揭示可能介导卵巢加速衰老的基因表达差异，我们对3周龄（青春期前）TAF4b敲除（TAF4b-null）与野生型（wild-type）小鼠的卵巢开展了全基因组表达谱对比分析。我们按照前述方法提取了9只3周龄小鼠的总RNA，其中野生型（wild-type）、TAF4b杂合子（TAF4b-heterozygous）与TAF4b敲除（TAF4b-null）小鼠各3只。采用生物分析仪（Bioanalyzer）检测RNA样品质量，利用超微量核酸定量仪（Nanodrop）测定RNA浓度。每份RNA样品取300 ng，用于Affymetrix全转录义链靶标标记实验（Rev 3版本），随后将标记产物与GeneChip小鼠基因1.0 ST芯片进行杂交。共使用9张基因芯片，以实现每种基因型的生物学重复三次。采用Affymetrix Expression Console软件（版本1.1）对数据进行标准化处理，并通过RMA-Sketch算法计算信号强度。