ExtFig6A_RACE_SLC7A5_R2_LG342.tif

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, 51 bp, 27 bp or 15 bp insertions from the BCAP31 or SLC7A5 genes, and where indicated, with PR8 PA-X. RNA was extracted and used to run 5’ RACE with primers annealing to luciferase sequences. DNA bands were purified and sequenced to confirm their identities

将HEK293T ishXrn1细胞用多西环素处理3~4天以诱导Xrn1基因敲低，随后转染携带来自BCAP31基因或SLC7A5基因的99 bp、51 bp、27 bp或15 bp插入片段的荧光素酶报告基因；在有标注的实验组中，同时转染PR8 PA-X。提取RNA后，使用靶向荧光素酶序列的引物开展5'末端快速扩增（5' Rapid Amplification of cDNA Ends, 5' RACE）实验，对所得DNA条带进行纯化并测序以确认其序列身份。