Gene resistance to transcriptional reprogramming following nuclear transfer is directly mediated by multiple chromatin repressive pathways. Supplementary Files (Replicate 3)

Apotome Images (raw data) of 2-cell fertilized mouse embryos with USP21 mRNA injection (inj) or not (T). Embryos are injected at fertilization with mRNA encoding USP21 and subsequently fixed for immunolabelling at the 2-cell stage with anti-H2AK119ub primary antibody (Cell Signaling #8240S + anti-rabbit Cy3-conjugated secondary antibody in red) and DAPI DNA conterstaining (in blue). A total number of 5 replicates were performed on 0140207, 20140211, 20140327, 20140505 and 20140506 (YYYY-MM-DD). In the three first experiments (Replicates 1/2/3), an internal microinjection reporter (HA-tag in green) was also included. In the last two replicates (20140505 and 20140506), co-detection of RING1B protein (Active Motif #39663 + anti-mouse FITC-conjugated secondary antibody in green) was included. Images file format: .zvi

本数据集包含经USP21 mRNA注射（inj组）或未注射（对照组T组）的2细胞期受精小鼠胚胎的原始落射光学切片（Apotome）图像。实验流程为：在受精阶段向胚胎注射编码USP21的mRNA，随后于2细胞期固定胚胎，使用抗H2AK119ub一抗（Cell Signaling #8240S）与Cy3偶联的抗兔二抗（荧光信号呈红色）进行免疫荧光标记，同时采用DAPI进行DNA复染（荧光信号呈蓝色）。本实验共完成5次生物学重复，实验日期分别为2014-02-07、2014-02-11、2014-03-27、2014-05-05及2014-05-06（日期格式为YYYY-MM-DD）。在前3次重复实验（重复1、2、3）中，同时加入了内部微注射报告基因HA标签（HA-tag，荧光信号呈绿色）。在后2次重复实验（2014-05-05及2014-05-06）中，额外增设了RING1B蛋白的共检测环节：使用抗RING1B一抗（Active Motif #39663）与FITC偶联的抗鼠二抗（荧光信号呈绿色）进行标记。本数据集的图像文件格式为.zvi。