图1:scaRNA2定位在DNA损伤和损失的RNA会导致DNA双链断裂的积累和基因组的不稳定。scaRNA2的结构示意图,说明GU-rich地区C / D域和U2核内小rna反义地区。smFISH探针结合位点,qPCR底漆,GapmeRs和一份循环插入网站也显示。b scaRNA2的亚细胞分布在未经处理或辐照U2OS细胞由qPCR决定。显示的值意味着±SD, n = 3。除非另有说明,所有n = 3指三个生理上独立的实验。Ns(不重要),由单向方差分析和双边Dunnett多个对比测试。c smRNA鱼卡哈尔scaRNA2和疣状的身体标记coilin U2OS细胞表达scaRNA2内生或过表达时24小时(n = 3)。核与DAPI染色在所有免疫荧光实验。d smRNA鱼scaRNA2和DNA损伤的免疫染色在激光标记γH2AX micro-irradiated(5分钟恢复)U2OS细胞overexpressing scaRNA2 24 h (n = 3)。 smRNA FISH for U2 snRNA was performed under the same conditions but without overexpression of scaRNA2. e smFISH of scaRNA2 in U2OS FokI cells transfected with a scaRNA2 plasmid for 24 h and treated with Shield and 4-OHT for an additional 4 h (n = 3). Immunostaining of γH2AX was performed under the same conditions but without overexpression of scaRNA2. f Immunostaining of γH2AX in irradiated (2 Gy, 1 or 24 h recovery) U2OS cells depleted or not of scaRNA2 for 48 h. The graph below shows the percentage of 100–200 cells (means ± SD, n = 3) whose nuclei contained > 10 γH2AX foci, **p < 0.01, ns (not significant) as determined by one-way ANOVA and two-sided Dunnett’s multiple comparisons test. The images depict a representative cell exhibiting the potential change in phenotype. g Immunostaining of γH2AX in untreated or irradiated (2 Gy, 1 or 24 h recovery) U2OS scaRNA2 WT or KO cells. The graph below shows the percentage of 100–200 cells (means ± SD, n = 3) whose nuclei contained more than 10 γH2AX foci, **p < 0.01, ns (not significant) as determined by unpaired two-tailed t-test. h DAPI staining in U2OS scaRNA2 WT or KO cells. The graph below shows the percentage of 100–200 cells (means ± SD, n = 3) that contained a micronucleus (indicated by a white arrowhead in the representative image), **p < 0.01 as determined by unpaired two-tailed t-test. Source data are provided as a Source Data file. Credit:DOI: 10.1038/s41467-022-28646-5
