Supporting data for “The Function of LEM-3 Endonuclease in Regulating Cell Fate Determination”

This project studies the effects of LEM-3 on cell fate determination in the nematode Caenorhabditis elegans. The study was mostly performed in the touch receptor neurons (TRNs) of C.elegans. There are six TRNs in C.elegans, including ALML, ALMR, PLML, PLMR, AVM, and PVM. We knocked out the lem-3 gene in C. elegans, labeled various neurons with fluorescent proteins driven by specific gene promoters, and observed the effects of lem-3 knockout on neuronal cell fate. We also used various biochemical methods to manipulate biological processes in C. elegans, such as using drugs to regulate the activity of topoisomerases to block the DNA replication process. One feature of the cell fate change caused by abnormal lem-3 is that its absence causes the formation of intercellular channels between neurons and sister neurons. We evaluated the degree to which cell fate was affected in different situations by quantifying the penetrance of intercellular canals.Figure 3C, 3D, 12C, 14B, 21: penetrance of intercellular canals in different neuronal subtypes; Figure 7E: percentage of independent and mixed transcription of genes in different types of syncytia; Figure 9B: length of intercellular canals at different time points during C.elegans development; Figure 10B, 10C: intensity of fluorescent signals in different neuronal subtypes; Figure 12B: percentage of nuclear membrane connections in syncytia; Figure 15C: penetrance of intercellular canals in AVM neurons under different experimental conditions. Figure 16A: percentage of different syncytia types; Figure 17E: Ratio of different cell fate-affecting outcomes.

本研究项目旨在探讨 LEM-3 对秀丽隐杆线虫 Caenorhabditis elegans 细胞命运决定的影响。研究主要在 C.elegans 的触觉受体神经元（TRNs）中开展。C.elegans 中共有六种 TRNs，包括 ALML、ALMR、PLML、PLMR、AVM 和 PVM。研究人员通过敲除 C.elegans 中的 lem-3 基因，利用特定基因启动子驱动的荧光蛋白标记各种神经元，并观察 lem-3 基因敲除对神经元细胞命运的影响。此外，研究者还运用多种生化方法操纵 C.elegans 中的生物过程，例如使用药物调节拓扑异构酶的活性以阻断 DNA 复制过程。由 lem-3 异常引起的细胞命运变化的一个特征是其缺失会导致神经元与邻近神经元之间形成细胞间隙通道。通过量化细胞间隙通道的侵润率，评估了在不同情况下细胞命运受影响的程度。图 3C、3D、12C、14B、21：不同神经元亚型中细胞间隙通道的侵润率；图 7E：不同类型合胞体中基因独立和混合转录的百分比；图 9B：C.elegans 发育不同时间点细胞间隙通道的长度；图 10B、10C：不同神经元亚型中荧光信号的强度；图 12B：合胞体中核膜连接的百分比；图 15C：不同实验条件下 AVM 神经元中细胞间隙通道的侵润率。图 16A：不同类型合胞体的百分比；图 17E：不同细胞命运影响结果的比率。