ExtFig3E_RACE_BCAP31_R1_LG274.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 insertions from the BCAP1, INSIG1, SLC7A5, STOML2, YKT6, and TUBA1B 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BCAP1、INSIG1、SLC7A5、STOML2、YKT6及TUBA1B基因99bp插入片段的荧光素酶报告基因载体，并在指定实验组中共转染PR8 PA-X。提取RNA后，使用与荧光素酶序列退火的引物开展5'末端快速扩增（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。