ExtFig5E_qPCR_R3_LG303.xlsx

HEK293T ishXrn1 cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with the luciferase reporters containing 99 bp insertions from the BCAP1, STOML2, YKT6, and TUBA1B genes, either the wildtype sequences or sequences with the GCUG mutated, as well as either empty vector or PR8 PA-X. RNA was extracted and levels were quantified by qRT-PCR

将诱导型Xrn1敲低HEK293T细胞（ishXrn1 cells）采用多西环素（doxycycline）处理3至4天以诱导Xrn1基因敲低，随后转染携带来自BCAP1、STOML2、YKT6及TUBA1B基因的99bp插入片段的荧光素酶报告基因（luciferase reporters），该插入片段分为野生型序列与GCUG突变序列两类；同时设置空载体或PR8 PA-X作为共转染组分。提取细胞总RNA后，通过实时定量逆转录聚合酶链反应（qRT-PCR）对RNA水平进行定量检测。