扩展数据图10:基因表达基因编辑tubuloids相比人类的肾脏和gydF4y2BaAVPR2gydF4y2Ba在tubuloids表达式。gydF4y2Ba
来自:gydF4y2Ba成年人肾脏瀑样源自CD24gydF4y2Ba+gydF4y2Ba细胞和代表着先进的成人多囊肾疾病的模型gydF4y2Ba
一个gydF4y2Ba,一般的维恩图分别表达下调的基因在塔尔集群TAL_2和人类ADPKD PT_4肾组织和人类健康的肾脏(供体活检)和基因编辑tubuloids控制tubuloids相比。gydF4y2BabgydF4y2Ba途径获得特异表达基因集富集和KEGG通路使用所有差异表达(DE)基因在人类ADPKD vs捐赠者活检PT-4和TAL-2细胞与细胞的tubuloids映射到PT-4和TAL-2使用交响乐。gydF4y2BacgydF4y2Ba,一般选择表达下调基因在人类ADPKD vs供体细胞活检和映射到PT-4 TAL-2gydF4y2BaPKDgydF4y2Ba−−/gydF4y2Ba与控制。gydF4y2BadgydF4y2Ba,结果CellTiter-Glo 3 d细胞生存能力分析的控制(EV),gydF4y2BaPKD1gydF4y2Ba−−/gydF4y2Ba和gydF4y2BaPKD2gydF4y2Ba−−/gydF4y2Batolvaptan tubuloids受到不同剂量。0.0显示0.2% DMSO tubuloids作为控制治疗。双向方差分析的因果Bonferroni n = 3意味着±s.e.m。DMSO控制细胞生存能力的差异不显著(0.0µM tolvaptan, 0.2% DMSO)相比,0.1 - 20µM tolvaptan EV tubuloids治疗,gydF4y2BaPKD1gydF4y2Ba−−/gydF4y2Ba,PKD2gydF4y2Ba−−/gydF4y2Batubuloids,gydF4y2BaPgydF4y2Ba> 0.9999;40µM tolvaptan vs DMSO控制,gydF4y2BaPgydF4y2Ba=gydF4y2Ba0.0846 EV tubuloids, * *gydF4y2BaPgydF4y2Ba= 0.0019gydF4y2BaPKD1gydF4y2Ba−−/gydF4y2Batubuloids, *gydF4y2BaPgydF4y2Ba=gydF4y2Ba0.0215gydF4y2BaPKD2gydF4y2Ba−−/gydF4y2Batubuloids。gydF4y2Bae-fgydF4y2Ba,DotplotsgydF4y2BaAVPR1A基因gydF4y2Ba和gydF4y2BaAVPR2gydF4y2Ba平均比例mRNA表达tubuloids(晚,好协议)(e)gydF4y2BaPKD1gydF4y2Ba−−/gydF4y2Ba以及gydF4y2BaPKD2gydF4y2Ba−−/gydF4y2Ba(f)。gydF4y2BaggydF4y2Ba,相对mRNA表达AVPR2 (V2R)摘要tubuloids相比tubuloids转导与慢病毒基因控制(EV)和tubuloids编辑lenti CRISPR / Cas9配对gydF4y2BaPKD1gydF4y2Ba或gydF4y2BaPKD2gydF4y2Ba。与因果Bonferroni 1-way-ANOVA n = 4意味着南达科他州±。,gydF4y2BaPKD1gydF4y2Ba−−/gydF4y2Batubuloids vs EV tubuloids, * * * *gydF4y2BaPgydF4y2Ba< 0.0001;gydF4y2BaPKD2gydF4y2Ba−−/gydF4y2Batubuloids vs EV tubuloids, * * *gydF4y2BaPgydF4y2Ba=gydF4y2Ba0.0006;gydF4y2BaPKD1gydF4y2Ba−−/gydF4y2Batubuloids vs正常tubuloids, * * * *gydF4y2BaPgydF4y2Ba< 0.0001;gydF4y2BaPKD2gydF4y2Ba−−/gydF4y2Batubuloids vs正常tubuloids, * * *gydF4y2BaPgydF4y2Ba=gydF4y2Ba0.0004;EV tubuloids vs tubuloids正常,gydF4y2BaPgydF4y2Ba> 0.9999。gydF4y2BahgydF4y2Ba人类ADPKD2 AVPR2代表原位杂交,肾组织切片。AVPR2 mRNA cyst-lining上皮细胞中检测到。规模50µm酒吧。gydF4y2Ba我gydF4y2Ba代表immunofluorescent LOH的染色标记SLC12A1 AVPR2集合管上皮细胞。规模50µm酒吧。gydF4y2BajgydF4y2Ba代表immunofluorescent LOH的染色标记SLC12A1和AVPR2 ADPKD囊肿衬里上皮细胞。规模50µm酒吧。在统计和再现性的详细信息,请参见方法。gydF4y2Ba
