Fig2E_18S-gel_R1-R3_LG379-374.sgd

<em>Xrn1 knock out A549 cells were infected with WT or PR8-PA(∆X), or mock infected for 16 hours before RNA extraction. 5’RACE was then performed using primers specific for STOML2,YKT6, TUBA1B, BCAP31 ~250-300 nucleotides (nt) downstream of the predicted cut sites. The PCR products were run on an agarose gel.</em> <em>HEK293T ishXrn1 cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with a luciferase reporter with or without the STOML2 99bp sequence insertion. RNA was extracted and analyzed by Northern blotting. Ethidium bromide pictures of gel showing rRNA are included.</em>

将Xrn1基因敲除的A549细胞分别以野生型（WT）或PR8-PA(∆X)病毒感染，或以模拟感染处理，培养16小时后提取RNA。随后使用针对STOML2、YKT6、TUBA1B、BCAP31基因，且位于预测切割位点下游250~300核苷酸（nt）的特异性引物开展5'末端快速扩增（5’RACE）实验，所得PCR产物经琼脂糖凝胶电泳分离。
将HEK293T ishXrn1细胞用多西环素处理3~4天以诱导Xrn1基因敲低，随后转染携带或不携带STOML2基因99bp序列插入的荧光素酶报告基因载体。提取RNA后通过Northern印迹杂交（Northern blotting）进行分析，同时附上溴化乙锭染色凝胶显示核糖体RNA（rRNA）的成像结果。