Fig2B_RACE_STOML2_R2_LG292_annotated.tiff

Xrn1 knock out A549 cells were infected with WT influenza A/Puerto Rico/8/1934 virus ("PR8") or PA-X deficient 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.将Xrn1基因敲除的A549细胞分别感染野生型甲型流感病毒A/Puerto Rico/8/1934（简称PR8）、PA-X缺陷型PR8-PA(ΔX)，或进行模拟感染；感染16小时后提取RNA。随后，针对预测剪切位点下游约250~300个核苷酸（nt）处的STOML2、YKT6、TUBA1B及BCAP31基因设计特异性引物，开展5'末端快速扩增（5' Rapid Amplification of cDNA Ends, 5'RACE）实验。将所得PCR产物进行琼脂糖凝胶电泳。