Expression data from E12.5 embryonic kidneys isolated from wild-type and Ret-deficient mice

To better understand the signaling and transcriptional events involved in the GDNF-independent emergence of the ureteric bud from the Wolffian duct, microarray expression analysis was performed on embryonic kidneys from wild-type and Ret-deficient mice. Microarray data was used to identify genes and gene networks involved in the GDNF-independent outgrowth of the ureteric bud. Whole embryonic kidneys from E12.5 Ret mutant and wild-type mice were isolated. Isolated kidneys were lysed and RNA was extracted with the Qiagen RNEasy Micro kit. The RNA was amplified using the NuGEn Ovation kit and hybridized to the Affymetrix GeneChip Mouse Whole Genome 430 2.0 microarray. Three biological replicates for Ret-knockout and wild-type kidneys were performed.为深入探究不依赖胶质细胞源性神经营养因子（Glial cell line-derived neurotrophic factor, GDNF）的输尿管芽（ureteric bud）从沃尔夫管（Wolffian duct）萌发过程中的信号转导与转录调控事件，本研究对野生型与Ret缺陷型小鼠的胚胎肾脏开展了基因表达微阵列分析。本研究利用该微阵列数据，筛选出参与不依赖GDNF的输尿管芽出芽生长过程的基因及基因调控网络。实验中分离了胚胎发育第12.5天（Embryonic day 12.5, E12.5）的Ret突变型与野生型小鼠完整胚胎肾脏。将分离得到的肾脏组织裂解后，使用Qiagen RNEasy Micro试剂盒提取总RNA。采用NuGEn Ovation试剂盒对RNA进行扩增，随后与Affymetrix GeneChip Mouse Whole Genome 430 2.0基因芯片进行杂交。本研究针对Ret基因敲除型与野生型胚胎肾脏各设置了3次生物学重复。