Identification of FOSL1 transcriptional targets in mutant KRAS lung adenocarcinoma cells. Homo sapiens

Mutant KRAS lung adenocarcinoma cells, H2009, were depleted of FOSL1 by a specific shRNA and its transcriptome was profiled Overall design: H2009 cells were infected with lentivirus carrying a FOSL1 or a control (GFP) shRNA for 2 days. Then, cells were selected for 3 days using 5ug/ml puromycin. After that, cells were counted and plated at a density of 600,000 cells/well (6-well plate). RNA was harvested on the following day using Trizol.

针对携带KRAS突变的肺腺癌细胞系H2009，采用特异性短发夹RNA（short hairpin RNA, shRNA）敲低FOSL1基因的表达，并对其转录组进行表达谱分析。总体实验设计：将H2009细胞感染携带FOSL1或对照（绿色荧光蛋白，green fluorescent protein, GFP）短发夹RNA的慢病毒，感染时长为2天；随后以5μg/mL嘌呤霉素筛选细胞3天；之后对细胞进行计数，以600,000个细胞/孔的密度接种于6孔板中；次日采用Trizol试剂提取总RNA。