mir - 142 - 3表达水平差- p在成纤维细胞和iPS。(a)表达了mir - 142 - 3 - p RT-qPCR在各种细胞。从表示细胞总RNA提取,并使用TaqMan RT-qPCR是微系统。U6 shRNA被用作控制。实验做了三次使用独立准备细胞,平均值和标准偏差。(b)的示意图表示反义- (-)mir - 142 - 3 - p EGFP表达质粒。LTR是用于驱动EGFP, U6启动子是用于驱动- mir - 142 - 3 - p。(c)过表达的影响- mir - 142 - 3 - p的表达水平内生mir - 142 - 3 - p在“诱导多能性”细胞。- mir - 142 - 3 - p / EGFP转染到ip或控制向量,,24小时后,mir - 142 - 3 - p水平iPS被RT-qPCR检查。数据表示为相对表达水平mir - 142 - 3 - p - mir - 142 - 3 - p / EGFP表达细胞控制向量表达的细胞。 Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers againstFgf5,Gata4,T brachyury,分别。值* 0.05 < 0.05和n。>,被学生的计算以及。