Sex-dependent and STAT5a-dependent Liver Gene Expression

A series of dual-channel gene expression profiles obtained using Agilent Mouse TOE 75k microarrays was used to examine the sex-dependent and STAT5a-dependent differences in gene expression in adult mouse liver. This series is comprised of 3 randomly chosen independent male and female wildtype mouse liver cDNA samples and 3 randomly chosen independent male and female STAT5a-deficient mouse liver cDNA samples, totalling 12 samples. The samples were paired randomly to generate 4 comparisons of M-WT:F-WT, M-WT:M-KO, F-KO:F-WT, F-KO:M-KO. Comparison of the set of sex-dependent genes with the set of genes responsive to the loss of STAT5a in females shows a small group of the sex-specific genes were also regulated by STAT5a in females. These results indicate a role for STAT5a in female gene expression to a lesser degree than that shown for STAT5b in males. Keywords: genetic knockout and sex response A Cy3-labeled cDNA sample is co-hybridized with a Cy5-labeled cDNA sample depending on the comparison being made. The samples are then dye-swapped and compared again on a second microarray chip. The two pairs are averaged together, the averages are normalized across the fluorescent reverse pairings, and the normalized ratio is reported along with the two separate intensities. This is replicated 3 times with independent samples of cDNA for each pair being compared. There are a total of 4 pairs of conditions being compared for a grand total of 12 samples.

本数据集采用安捷伦（Agilent）小鼠TOE 75k基因微阵列获取双通道基因表达谱，用于探究成年小鼠肝脏中基因表达的性别依赖型与信号转导与转录激活因子5a（STAT5a）依赖型差异。
本数据集包含3份随机选取的独立野生型雌雄小鼠肝脏互补DNA（cDNA）样本，以及3份随机选取的独立STAT5a缺陷型雌雄小鼠肝脏cDNA样本，总计12份样本。
样本经随机分组，形成4组比对：M-WT:F-WT、M-WT:M-KO、F-KO:F-WT及F-KO:M-KO。
将性别依赖型基因集合与雌性中响应STAT5a缺失的基因集合进行比对后发现，仅有少量性别特异性基因同时受雌性体内STAT5a的调控。上述结果表明，STAT5a在雌性小鼠基因表达调控中所发挥的作用，弱于其同源蛋白信号转导与转录激活因子5b（STAT5b）在雄性小鼠中的调控作用。
关键词：基因敲除与性别应答
根据待进行的比对组别，将经Cy3标记的cDNA样本与经Cy5标记的cDNA样本进行共杂交。随后进行染料互换实验，将互换后的样本在第二张基因芯片上再次进行杂交比对。对这两组杂交数据取平均值，随后针对荧光反转配对的样本进行归一化处理，最终报告归一化比值以及两组独立的荧光强度值。每组比对均使用独立的cDNA样本重复实验3次。总计有4组实验条件比对，对应全部12份样本。