Ath_Agilent_v2_Riechmann array expression profile AP1-GR ap1cal inflorescences - chx experiment

This experiment describes gene expression after the activation of APETALA1-GR, to study and identify AP1 target genes. We used a 35S:AP1-GR ap1 cal line to induce a synchronized response activating the AP1-GR fusion protein in ap1 cal inflorescence-like meristems through dexamethasone or dexamethasone+cycloheximide treatment. Tissue samples were collected at 3hrs after the treatment. The expression profiles of the individual samples were then analyzed by gene expression profiling using whole-genome oligonucleotide arrays (Agilent, custom-commercial). We treated inflorescences of 35S:AP1-GR ap1-1 cal-1 plants with a dexamethasone-containing or a mock solution, or with identical solutions that contained in addition 10 μM cycloheximide. Tissue was collected 3 hours after the treatment. Samples from each of the four biological replicates resulted in a set of four hybridization pairs: Mock vs. Dex, Mock vs. Chx, Mock vs. Dex+Chx, and Chx vs. Dex+Chx. Dye polarities were switched between biological replicates.本实验旨在研究并鉴定AP1（APETALA1）的靶基因，分析APETALA1-糖皮质激素受体融合蛋白（APETALA1-GR）激活后的基因表达特征。我们采用携带35S:AP1-GR的ap1 cal株系，通过地塞米松（dexamethasone, Dex）或地塞米松联合环己酰亚胺（cycloheximide, Chx）处理，在该株系的类花序分生组织中同步激活AP1-GR融合蛋白，于处理后3小时收集组织样本，随后采用全基因组寡核苷酸芯片（whole-genome oligonucleotide arrays，Agilent定制商业化产品）对各样本的表达谱进行分析。本实验以35S:AP1-GR ap1-1 cal-1植株的花序为材料，分别施加含地塞米松的溶液、空白对照溶液，以及额外添加10μM环己酰亚胺的同类型对照溶液，于处理后3小时采集组织样本。本实验设置4次生物学重复，共形成4组杂交比对组合：Mock vs. Dex、Mock vs. Chx、Mock vs. Dex+Chx及Chx vs. Dex+Chx。不同生物学重复间进行染料极性互换，以消除染料偏好性带来的实验偏差。