Fig4B_RACE_STOML2_R1_AI8.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 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细胞经多西环素（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)来源的PA-X表达质粒或空载体，以及携带STOML2或YKT6基因插入序列的荧光素酶报告基因质粒。插入片段大小范围为15 bp至99 bp。于转染后24小时收集RNA样品，采用结合荧光素酶序列的引物，通过5' RACE（5' Rapid Amplification of cDNA Ends）进行分析。