Expression data of the control and CITED1 OE ESCs. Mus musculus

Trophoblast lineages, as the precursor of placenta, are essential for post-implantation embryo survival. However, the regulatory networks for trophoblast development remains incompletely understood. Here, we identified CITED1 as a regulator to induce trophoblast-like differentiation from mESCs. Overexpression of CITED1 in ESCs prompted differentiation towards trophoblast accompanying with elevated expression of trophoblast marker genes. To evaluate the ability of CITED1 to induce trophoblast differentiation at a genome-wide scale, we compared the global transcriptional profiles between CITED1 overexpressing cells and control ESCs by Affymetrix microarray analysis at day 1 and day 2 after transfection. We used microarrays to identify genes affected by CITED1 overexpression in mouse ESCs. Overall design: E14T ESCs were transiently transfected with plasmids expressing CITED1 or empty vector pPy. 8 hours later, ESCs were selected by puromycin at the final concentration of 2 ug/mL, and then collected for RNA isolation. In total, 15 RNA samples were prepared from three independent biological triplicates. Microarray was carried out with the Affymetrix GeneChip® Mouse Genome 430 2.0 Arrays. The procedures including RNA extraction, cDNA synthesis, labeling, hybridization, washing, and scanning were conducted according to the standard Affymetrix protocol by the Shanghai Biotechnology Corporation.