Effect of butyrate and DZNep on hESC HSF-6

In order to recover nuclei with two active X chromosomes (class I), we developed a reprogramming strategy by supplementing hESC media with the small molecules sodium butyrate, and 3-deazaneplanocin A (DZNep). In order to determine how B+D affects global gene expression, we performed microarray analysis in triplicate in the HSF-6 (8) C and HSF-6 (8) B+D treated cultures. We also evaluated HSF-6 (S9) B+D in triplicate and identified no statistically significant changes in gene expression in HSF-6 (S9) B+D compared to HSF-6 (8) B+D treated cultures. This suggests that global transcriptional differences are more strongly modulated by presence or absence of B+D and not the percentage of class I, II or III nuclei. We performed gene expression profiling on hESC HSF-6 (8, S9) in absent (control) and presence of butyrate and DZNep. All cell were collected after 11 passages in absent and presence of butyrate and DZNep.

为获取携带两条活性X染色体的细胞核（I类），我们开发了一种重编程策略：在人类胚胎干细胞（human embryonic stem cell, hESC）培养基中添加小分子化合物丁酸钠与3-脱氮腺苷（3-deazaneplanocin A, DZNep）。为探明丁酸钠与DZNep联合处理（B+D）对全局基因表达的影响，我们对HSF-6 (8) 对照组与HSF-6 (8) B+D处理组的培养细胞开展了三次重复微阵列分析。此外，我们还对HSF-6 (S9) B+D处理组进行了三次重复分析，结果显示，与HSF-6 (8) B+D处理组相比，HSF-6 (S9) B+D处理组的基因表达无统计学显著差异。这表明全局转录组差异主要受B+D处理与否调控，而非I、II或III类细胞核的占比。我们对HSF-6 (8, S9) 人类胚胎干细胞进行了基因表达谱分析，实验分为未处理对照组与丁酸钠、DZNep处理组。所有细胞均在经11次传代培养（分别处于未处理与丁酸钠、DZNep处理的培养环境中）后收集。