Elevating Sox2 levels deleteriously affects the growth of glioblastoma and medulloblastoma cells.. Homo sapiens

Induction of the transcription factor Sox2 from a doxycycline-inducible promoter in iSox2-DAOY medulloblastoma cells. Affymetrix microarrays were used to characterize the gene expression profile of iSox2-DAOY cells in the absence and presence of doxycycline. Overall design: Human DAOY medulloblastoma cells were engineered to express epitope tagged Sox2 under the control of a doxycycline inducible promoter, to produce iSox2-DAOY cells. iSox2-DAOY cells were FACS sorted into CD133 low and high populations. The CD133 low population was cultured in the absence and presence of 0.5 µg/mL doxycycline for 24 hours. RNA was extracted from cells cultured without and with doxycycline, and was used for microarray analysis.

在iSox2-DAOY髓母细胞瘤细胞中，通过多西环素诱导型启动子诱导转录因子Sox2的表达。采用Affymetrix微阵列（Affymetrix microarrays）分析iSox2-DAOY细胞在有、无多西环素处理条件下的基因表达谱。
整体实验设计：将人源DAOY髓母细胞瘤细胞进行基因工程改造，使其在多西环素诱导型启动子的调控下表达表位标签标记的Sox2，从而构建得到iSox2-DAOY细胞。通过荧光激活细胞分选（FACS）将iSox2-DAOY细胞分为CD133低表达与高表达两个亚群。取CD133低表达亚群，分别在不含以及含有0.5 μg/mL多西环素的培养基中培养24小时。从经多西环素处理及未处理的细胞中提取RNA，用于微阵列分析。