Gene expression profiles of Fir1 (RNAi) animals

The goal of this study was to identify Fir1 downstream genes that were required for local proliferation. Fir1(RNAi) animals displayed reduced local proliferation near the wound,while the proliferation far from the wound was not decreased after Fir1 RNAi. The regenerating pieces following Fir1 RNAi were expected to only show decreased levels of the genes near the wound relative to control RNAi animals. Animals were administrated by Fir1 dsRNA for 8 days, amputated after pharynx. 48 hours later, RNA from the region adjacent to the wound(part1) and the region far from the wound(part2) in Fir1(RNAi) tail pieces was harvested with Trizol. Two biological replicates were used. Then the RNA was hybridized on a microarray with RNA from control RNAi animals.

本研究旨在鉴定介导局部增殖所必需的Fir1下游基因。经Fir1 RNA干扰（RNA interference, RNAi）处理的动物，其伤口邻近区域的局部增殖水平显著降低，而远离伤口的增殖在Fir1 RNA干扰后并未出现下降。研究预期，与对照RNA干扰组动物相比，Fir1 RNA干扰后的再生片段仅在伤口附近区域呈现基因表达水平的下调。实验中，受试动物经Fir1双链RNA（double-stranded RNA, dsRNA）处理8天后，于咽部实施截肢操作。术后48小时，采用TRIzol试剂提取Fir1 RNA干扰组尾部片段中伤口邻近区域（区域1）与远离伤口区域（区域2）的总RNA。本实验设置两个生物学重复。随后，将提取的总RNA与对照RNA干扰组动物的RNA进行基因芯片（microarray）杂交。