IRF4-dependent division-coupled reprogramming events [IRF4cKO.RNAseq]

To assess the timing and scope of IRF4–dependent reprogramming in vivo, CTV-labeled IRF4–sufficient and –deficient B cells were transferred into mMT hosts. One day later, hosts were challenged with 50 mg LPS. Three days post-LPS challenge, transferred cells were recovered and sorted from divisions 0, 1, 3, 4, 5, and 6 as determined by CTV dilution for RNA–seq. Overall design: RNA-seq on control and IRF4-deficient B cells at discrete divisions为体内评估干扰素调节因子4（IRF4）依赖性重编程的时序与范围，将经细胞追踪染料CTV（CellTrace Violet）标记的IRF4充足型与IRF4缺陷型B细胞转移至mMT缺陷宿主。造模后1日，以50 mg脂多糖（LPS）刺激宿主。脂多糖刺激后第3日，回收移植细胞，基于CTV荧光稀释程度，从第0、1、3、4、5、6代增殖细胞中分选样本，用于RNA测序（RNA-seq）。实验整体设计：对不同增殖代数的对照组与IRF4缺陷型B细胞开展RNA-seq测序。