Fig4B_RACE_YKT6_R1_AI9_annotated.tiff

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 inserted sequence from the STOML2 or YKT6 gene. Insertion size ranges from 15 bp to 99 bp. 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，随后转染来自甲型流感病毒A/波多黎各/8/1984（H1N1）的PA-X蛋白或空载体（vector），以及插入了STOML2或YKT6基因序列的荧光素酶报告基因（luciferase reporters）。插入片段的长度范围为15 bp至99 bp。24小时后收集RNA，采用结合荧光素酶序列的引物通过5'末端快速扩增（5' RACE）技术完成分析。