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Supporting data for "Characterization of epigenetic regulation during early trophoblast differentiation human expanded potential stem cells"

posted on 2023-09-26, 06:05 authored by Wen HuangWen Huang

This study used embryonic stem cells, namely human expanded potential stem cells (EPSCs), to investigate the epigenetic regulation during human preimplantation development, and the main focus is the trophoblast differentiation. Several experimental techniques were used in this study, including but not limited to RT-qPCR, western blotting, immunocytochemical staining, and fluorescent signal reading. In addition, the published datasets were analyzed in this study to identify the novel target genes regulated by DNA methylation. The function of the target genes was then investigated by CRISPR/Cas9-mediated gene knockout. The roles of Argonaute 2/microRNA complexes during trophoblast differentiation were also determined using CRISPR/Cas9 approach.

The major part of the raw data in the dataset is the gene expression analysis using RT-qPCR. Each value represents the gene's relative level in this group compared to the control group. The western blotting was first analyzed using ImageJ, and the intensities of protein bands were recorded in this dataset. Similarly, the data were normalized to the control group. In addition, the fluorescent signal of mCherry was detected by the microplate reader and the exact values were recorded. The DNA methylation level at the gene promoter was detected by Bisulfite sequencing, and the average promoter methylation level was calculated as the overall average methylation level of each CpG site in the gene promoter. The analysis of the published data was conducted using R Studio. The final output data were included in this dataset.