Fig3F_YKT6_ExtFig5F_TUBA1B_RACE_R2_LG302_annotated.tif

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 YKT6 or TUBA1B gene, 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.

HEK293T ishXrn1细胞经强力霉素（doxycycline）处理3~4天以诱导Xrn1基因敲低，随后分别转染甲型流感病毒A/Puerto Rico/8/1984（H1N1）株来源的PA-X蛋白表达载体或空载体，以及携带源自YKT6或TUBA1B基因的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与携带GCTG→TAGC突变的两种类型。转染24小时后收集细胞RNA，采用结合荧光素酶序列的引物通过5' 快速扩增cDNA末端（Rapid Amplification of cDNA Ends, 5' RACE）进行分析。