全基因组TF筛选确定Atf7ip, JunB, Sp7和Zfp207 (ZNF207)是细胞命运重编程的障碍。心脏重编程实验原理示意图。b描绘全基因组TF筛选结果的火山图。x轴为Myh6-EGFP +阳性归一化至siccontrol的百分比。Y轴表示与siccontrol相比P值的−log。筛选试验重复四次。c对前20个命中数的验证确定了8个具有确认活性的siRNAs。学生的学习任务:* * * p < 0.05, p < 0.01, * * * p < 0.001, * * * * p < 0.0001。d MGT诱导后第3天，转染siAtf7ip-和siccontrol -的imgt - mef的代表性图像。Myh6-eGFP为绿色，细胞核用DAPI染色(右上方的蓝色插图)。 e Schematic of the combinatorial screening approach. A total of 255 combinations was tested, each in quadruplicate. f Volcano plot depicting genome-wide combinatorial screen results. Data were normalized and compared to the most potent single hit (siAtf7ip). 1–4 indicates top combinations that were significantly more potent than siAtf7ip alone (p < 0.05). siAtf7ip, siJunb, siSp7, and siZfp207 (siAJSZ) are shown in red. g Visualization of top 4 siRNA combinations in a histogram plot as in (c). h Representative images of iMGT-MEFs transfected with siAJSZ (siAtf7ip, siJunb, siSp7, and siZfp207) 3 days after MGT induction. n = 4 per condition for all data in this figure. Groups were compared using two-tailed unpaired analysis. Data in the figure are presented as mean values ± standard deviation. Scale bars: 50 µm. a, e Schematics are modified from Cunningham, T. J. et al. Id genes are essential for early heart formation. Genes & development, https://doi.org/10.1101/gad.300400.117 (2017). - CC-BY 4.0. Credit:自然通讯(2023)。DOI: 10.1038 / s41467 - 023 - 37256 - 8
