Figure_4_replicates

(B, F, G) HEK293T ishXrn1cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with luciferase reporters containing insertions of the indicated lengths from the YKT6 or STOML2 genes (B). In F and G, constructs also carry mutations that disrupt the hairpin structure and restore with a different sequence. PR8 PA-X or an empty vector were also co-transfected. RNA was extracted to run 5’ RACE. DNA bands were purified and sequenced to confirm their identities. R# --> replicate number; AI#/LG### --> experiment number. .sgd are the original source files acquired with the Syngene Imager.(B、F、G组) 将HEK293T ishXrn1细胞经多西环素处理3~4天以诱导Xrn1基因敲低，随后转染携带YKT6或STOML2基因指定长度插入片段的荧光素酶报告基因载体（对应图B）。在F、G组实验中，所用重组载体还携带可破坏茎环结构并通过不同序列进行修复的突变位点。同时共转染PR8 PA-X或空载体。提取总RNA后开展5'末端快速扩增（5' RACE）实验。对DNA条带进行纯化并测序，以确认其序列身份。其中R#代表重复次数；AI#/LG###代表实验编号。后缀为.sgd的文件为通过Syngene成像仪获取的原始源文件。