Fig3F_STOML2_ExtFig5F_BCAP31_RACE_R1_LG301.sgd

HEK293T ishXrn1 cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with PA-X from the influenza A/Puerto Rico/8/1984 (H1N1) or vector, and luciferase reporters with a 99 bp inserted sequences from the STOML2 or BCAP31 genes, either wild-type or containing a GCTG --> TAGC mutation. RNA was collected 24 hrs later and analyze by 5' RACE using a primer that binds to luciferase sequences.

将诱导型Xrn1敲减HEK293T细胞（HEK293T ishXrn1 cells）经多西环素处理3-4天以诱导Xrn1敲减，随后分别转染甲型流感病毒A/波多黎各/8/1984（H1N1）亚型的PA-X表达质粒或空载体，以及携带STOML2或BCAP31基因99 bp插入序列的荧光素酶报告基因质粒；该插入序列分为野生型和携带GCTG→TAGC突变的两种类型。转染24小时后收集RNA，采用结合荧光素酶序列的引物，通过5' cDNA末端快速扩增（5' RACE, 5'-Rapid Amplification of cDNA Ends）技术进行分析。