ExtFig6A_RACE_SLC7A5_R2_LG342.sgd

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细胞以多西环素（doxycycline）处理3~4天以诱导Xrn1基因敲低；随后转染携带来自BCAP31或SLC7A5基因的99 bp、51 bp、27 bp或15 bp插入片段的荧光素酶（luciferase）报告基因载体，且在标注的实验组中共同转染PR8 PA-X。提取细胞总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' RACE）实验，纯化所得DNA条带并进行测序，以验证其序列身份。