POGZ耗尽导致ESC失去自我更新。A(上)描绘gRNA靶向小鼠Pogz基因外显子2位点的漫画;(下)Pogz基因突变等位基因1 (Mut1)的基因分型。B qRT-PCR显示突变ESCs中Pogz表达减少。C对照POGZ和POGZ−/−ESCs的Western blot分析。D OCT4和POGZ双IF染色显示OCT4水平在早期传代突变ESCs中没有明显改变。E细胞生长治愈显示，与对照组相比，Pogz−/−和Pogz+/−ESCs的增殖缺陷。第12代的Mut1 ESCs用于检测。F对照和晚期传代Pogz−/−ESCs形态的代表图像(传代号为18)。白色箭头指向扁平的分化细胞。 G Alkaline phosphatase staining of control and Pogz−/− ESCs. White arrows pointing to cells with reduced AP activities. H The mRNA expression levels of representative pluripotency-related, mesodermal, neuroectodermal, endodermal genes in control and Pogz−/− ESCs. I IF staining of GATA6 showing that GATA6 was abnormally expressed in Mut1 Pogz−/− ESCs. Restoring POGZ rescued the abnormal expression of Gata6. J WB showing that OCT4 levels were reduced in Pogz−/− ESCs, which can be reversed by reintroducing FLAG-POGZ. Bar: 25 μm. K Volcano plot showing the up- and down-regulated genes in the control and early passage Pogz−/− ESCs. The RNA-seq experiments were repeated two times. L GO analysis of up-regulated DEGs. M GO analysis of down-regulated DEGs. All WB and IF were repeated at least two times. qRT-PCR was repeated three times. Credit:分子自闭症(2022)。DOI: 10.1186 / s13229 - 022 - 00502 - 9
