Fig2F_YKT6_ExtFig4C_TUBA1B_RACE_R1_LG295.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) ("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（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，简称"PR8"）、A/Perth/16/2009（H3N2，简称"Perth"）毒株的PA-X编码序列或空载体，同时转染带有或不带有STOML2、YKT6、BCAP31及TUBA1B基因99bp插入片段的荧光素酶（luciferase）报告基因载体。转染24小时后收集总RNA，采用结合荧光素酶序列的引物，通过5'末端快速扩增（5' Rapid Amplification of cDNA Ends, 5' RACE）技术进行分析。（Cal09构建体存在实验缺陷，该批次样本已在独立实验中重新完成。）