Ath29 congenic mice - gene expression in aorta

A congenic mouse line was constructed by introgressing a C3H chromosome 9 region harboring Ath29 into the C57BL/6 apoE-deficient background. RNA was extracted from aorta using a QIAGEN kit . Total RNA was pooled in an equal amount from 3 mice for each group. Standard Affymetrix procedures were performed using 8ug of total RNA. Microarrays were used to detect gene expression in the aorta of Ath29 congenic mice and C57BL/6 apoE-deficient mouse strains fed a chow or western diet. 4 groups of mice were studied: C57BL/6 apoE-/- control mice and Ath29 congenic mice fed a chow or Western diet. After fed a Western diet for 2 weeks, aorta was harvested and total RNA prepared.本研究构建了同源导入系小鼠（congenic mouse line）：将携带Ath29基因座的C3H小鼠9号染色体区域回交导入至C57BL/6背景的载脂蛋白E（apoE）敲除小鼠基因组中。采用QIAGEN试剂盒从主动脉中提取总RNA，每组取3只小鼠的总RNA进行等量混合。使用8 μg总RNA，按照标准Affymetrix实验流程完成样本制备。采用基因芯片（microarray）技术，检测喂食普通饲料（chow diet）或西方高脂饮食（western diet）的Ath29同源导入系小鼠，以及C57BL/6背景apoE敲除（apoE-/-）小鼠的主动脉基因表达水平。本研究共设置4组实验小鼠：分别为喂食普通饲料、西方高脂饮食的C57BL/6背景apoE敲除（apoE-/-）对照小鼠，以及喂食普通饲料、西方高脂饮食的Ath29同源导入系小鼠。喂食西方高脂饮食的小鼠在造模2周后采集主动脉并制备总RNA。