Supplementary Material for: Jagged1 is Clinically Prognostic and Promotes Invasion of Glioma-Initiating Cells by Activating NF-κB(p65) Signaling

<b><i>Background/Aims:</i></b> Jagged1 is the ligands of the Notch signaling and has been shown to promote glioma-initiating cells (GICs) in glioblastoma. The role of Jagged1 in GICs invasion and underlying molecular mechanisms remain unclear. <b><i>Methods:</i></b> Survival data from R2 genomics analysis, the Cancer Genome Atlas (TCGA), the Chinese Glioma Genome Atlas (CGGA) and visualization platform database were used to evaluate the effects of Jagged1 on overall patient survival. we investigated Jagged1 induced the GICs cells’ invasion by matrix degradation assays and Transwell cell invasion assays <i>in vitro</i>, then we further explored the underlying molecular mechanisms using Co-immunoprecipitation (co-IP) analysis. <b><i>Results:</i></b> High expression of Jagged1 in human glioma was associated with poor survival. Clinical data analysis showed that the Jagged1 was positively correlated with NF-κB(p65). Jagged1-induced invasion of GICs cells through activation of NF-κB(p65) pathway. <i>In vivo</i>, knockdown of Jagged1 could suppress the tumorigenicity of GICs cells through NF-κB(p65) signaling. <b><i>Conclusion:</i></b> Insights gained from these findings suggest that Jagged1 plays an important oncogenic role in GICs malignancy by activation of NF-κB(p65) signaling, and Jagged1 could be employed as an effective therapeutic target for GICs.

<b><i>背景与目的：</i></b> Jagged1是Notch信号通路的配体，已有研究证实其可促进胶质母细胞瘤中的胶质瘤起始细胞（glioma-initiating cells, GICs）。目前Jagged1在GICs侵袭中的作用及其潜在分子机制尚不明确。<b><i>方法：</i></b> 本研究利用R2基因组分析、癌症基因组图谱（The Cancer Genome Atlas, TCGA）、中国胶质瘤基因组图谱（Chinese Glioma Genome Atlas, CGGA）及可视化平台数据库的生存数据，评估Jagged1对患者总生存期的影响。本研究通过基质降解实验与Transwell细胞侵袭实验体外探究Jagged1诱导的GICs侵袭行为，并进一步采用免疫共沉淀（Co-immunoprecipitation, co-IP）分析揭示其潜在分子机制。<b><i>结果：</i></b> 人类胶质瘤中Jagged1的高表达与不良预后相关。临床数据分析显示，Jagged1与NF-κB(p65)呈正相关。Jagged1通过激活NF-κB(p65)通路诱导GICs侵袭。体内（in vivo）实验表明，敲低Jagged1可通过NF-κB(p65)信号通路抑制GICs的成瘤能力。<b><i>结论：</i></b> 本研究结果提示，Jagged1可通过激活NF-κB(p65)信号通路在GICs恶性表型中发挥重要的致癌作用，且Jagged1可作为GICs的有效治疗靶点。