Fig3F_YKT6_ExtFig5F_TUBA1B_RACE_R3_LG303.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 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.

将HEK293T ishXrn1细胞用多西环素（doxycycline）处理3~4天以诱导Xrn1基因敲低，随后转染甲型流感病毒A/Puerto Rico/8/1984（H1N1）来源的PA-X或空载体，同时转染携带YKT6或TUBA1B基因99 bp插入序列的荧光素酶报告基因（luciferase reporters）；该插入序列分为野生型与携带GCTG→TAGC突变的两种。于转染后24小时收集RNA，使用结合荧光素酶序列的引物通过5'快速扩增cDNA末端（5' RACE）完成分析。