Gene regulation by Ffr2 in the conjunctival epithelium

Wild type or Fgfr2 fx/fx; Le-Cre-positive (Fgfr2-/-) conjunctival epithelium of E14.5 mouse embryos was laser micodissected from three embryos of each genotype. Total RNA was purified and reverse transcribed and amplified using a NuGEN kit. cDNA was biotinylated and hybridized to Illumina MouseWG-6 v2.0 bead arrays. Three wild type and three Ffr2 knockout embryos were used. cDNA obtained from each was used for microarray analysis. Statistical analysis was performed using Illumina Genome Studio software.本研究从胚胎发育第14.5天（E14.5）的小鼠胚胎中，分别从每种基因型的3个胚胎中通过激光显微切割（laser microdissection）获取野生型或Fgfr2 fx/fx; Le-Cre阳性（即Fgfr2基因敲除型，Fgfr2-/-）的结膜上皮组织；随后纯化总RNA，采用NuGEN试剂盒进行反转录与扩增；将生物素标记的互补脱氧核糖核酸（complementary DNA, cDNA）与Illumina MouseWG-6 v2.0微珠芯片进行杂交。本实验共纳入3个野生型胚胎与3个Fgfr2基因敲除胚胎，每份样本的cDNA均用于芯片分析，统计分析通过Illumina Genome Studio软件完成。