Fig2F_STOML2_ExtFig4C_BCAP31_RACE_R2_LG297.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) ("PR8") or A/Perth/16/2009 (H3N2) ("Perth") strains or vector, and luciferase reporters with or without 99 bp inserted sequences from the STOML2, YKT6, BCAP31 or TUBA1B genes. RNA was collected 24 hrs later and analyze by 5' RACE using a primer that binds to luciferase sequences. (Cal09 construct had an issue and these samples were re-done in separate experiments).将HEK293T ishXrn1细胞用多西环素处理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)（简称"PR8"）、A/Perth/16/2009 (H3N2)（简称"Perth"）毒株的PA-X蛋白，或空载体，同时转染携带或不携带STOML2、YKT6、BCAP31或TUBA1B基因99 bp插入序列的荧光素酶报告基因。于转染后24小时收集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）技术进行分析。(Cal09构建体存在实验缺陷，该批次样本已通过独立实验重新完成)