UTX condensation underlies its tumor suppressive activity

We investigated whether UTX condensates regulate local H3K4me1 in the genome by ChIP-seq of THP-1 cells expressing empty vector (control), UTX WT, ?cIDR, UTX-eIFIDR, or ?TPR. To determine the effect of UTX condensation on high-order chromatin interactions or looping, we performed H3K4me3 HiChIP in duplicates to generate high-resolution contact maps around the active and poised transcription start sites (TSSs) in THP-1 cells expressing empty vector (control), UTX WT, ?cIDR, or UTX-eIFIDR. Overall design: ChIP was performed and libraries were constructed and sequenced with 20 million THP-1 cells expressing empty vector (control), UTX WT, ?cIDR, UTX-eIFIDR, or ?TPR. HiChIP experiments were performed with 2.5 million THP-1 cells expressing empty vector (control), UTX WT, ?cIDR, UTX-eIFIDR, or ?TPR using the Arima-HiC+ kit according to its recommended protocol.

我们通过对表达空载体（empty vector，对照组）、野生型UTX（UTX WT）、ΔcIDR、UTX-eIFIDR或ΔTPR的THP-1细胞开展染色质免疫共沉淀测序（ChIP-seq），探究UTX凝聚体（UTX condensates）是否可调控基因组内的局部组蛋白H3赖氨酸4单甲基化（H3K4me1）水平。为明确UTX凝聚对高阶染色质互作或染色质环形成的影响，我们对表达空载体（empty vector，对照组）、UTX WT、ΔcIDR或UTX-eIFIDR的THP-1细胞中活性与预备态转录起始位点（transcription start sites, TSSs）附近区域进行了生物学重复的H3K4me3 HiChIP测序，以生成高分辨率染色质接触图谱。整体实验设计：分别收集2000万株表达上述各载体的THP-1细胞，开展染色质免疫共沉淀实验、构建文库并完成测序。同时，采用Arima-HiC+试剂盒（Arima-HiC+ kit）并遵循其推荐实验流程，分别收集250万株上述转染细胞进行重复HiChIP实验。