Transcriptome of GATA3 knocked-down and normal keratinocytes after 1 cGy X irradiation

To characterize the consequences of the GATA3 silencing on the transcriptome of X-rays irradiated keratinocytes. Oligonuclotide microarrays were used to assess transcriptional changes over a time-course between 3 and 72h post-irradiation in a GATA3 knocked-down background (shGATA3) or in a normal background (ShSCR). Primary human keratinocytes were cultured in a semi-defined medium and then infected either with shGATA3 or with ShSCR lentiviral particles . Cells were then cultured 5 to 8 days up to confluence and then submitted to an 1 cGy dose of X-rays. Total RNA was extracted 4, 24, 48 or 72 hours after irradiation. After RNA amplification and labeling, gene profiling was performed using oligonucleotide microarrays (26 068 probes) to compare 1 cGy irradiated cells to sham-irradiated cells at individual times. Dye-swap hybridization were performed. Slides were scanned with a Genepix 4000 microarray scanner (Axon Instruments, Molecular devices, Sunnyvale, CA). For each hybridized spot, the Cy3 and Cy5 fluorescence values were obtained by using Genepix Pro 4.0 software (Axon Instruments) and were saved as a result file

本数据集旨在解析GATA3基因沉默对X射线照射角质形成细胞转录组的影响。研究采用寡核苷酸微阵列（oligonucleotide microarray），在GATA3基因敲低（shGATA3）或正常对照（ShSCR）背景下，检测X射线照射后3至72小时时间进程内的转录水平变化。将原代人角质形成细胞培养于半限定培养基中，随后分别感染shGATA3或ShSCR慢病毒颗粒。待细胞培养5至8天至汇合状态后，给予1厘戈瑞（cGy）剂量的X射线照射。分别于照射后4、24、48及72小时提取总RNA。完成RNA扩增与标记后，采用包含26068个探针的寡核苷酸微阵列进行基因表达谱分析，在各个时间点分别比较1厘戈瑞照射组与假照射组的细胞样本，并开展染料交换杂交实验。使用Genepix 4000微阵列扫描仪（Axon Instruments, Molecular Devices, 加利福尼亚州桑尼维尔）对芯片进行扫描；通过Genepix Pro 4.0软件（Axon Instruments）获取每个杂交斑点的Cy3与Cy5荧光强度值，并保存为结果文件。